6-(5-membered heteroaryl)isoquinolin-3-yl-(5-membered heteroaryl) carboxamides and preparation and use thereof

ABSTRACT

Isoquinoline compounds for treating various diseases and pathologies are disclosed. More particularly, the present disclosure concerns the use of an isoquinoline compound or analogs thereof, in the treatment of disorders characterized by the activation of Wnt pathway signaling (e.g., cancer, abnormal cellular proliferation, angiogenesis, Alzheimer&#39;s disease, lung disease, inflammation, auto-immune diseases and osteoarthritis), the modulation of cellular events mediated by Wnt pathway signaling, as well as neurological conditions/disorders/diseases linked to overexpression of DYRK1A.

RELATED APPLICATION

The application is a continuation application of U.S. application Ser.No. 16/172,589, filed on Oct. 26, 2018 which claims benefit of U.S.Provisional Application No. 62/578,370, filed Oct. 27, 2017, both ofwhich are incorporated herein by reference in its entirety.

BACKGROUND Technical Field

This disclosure relates to inhibitors of one or more proteins in the Wntpathway, including inhibitors of one or more Wnt proteins, andcompositions comprising the same. More particularly, it concerns the useof an isoquinoline compound or salts or analogs thereof, in thetreatment of disorders characterized by the activation of Wnt pathwaysignaling (e.g., cancer, abnormal cellular proliferation, angiogenesis,Alzheimer's disease, lung disease, inflammation, auto-immune diseasesfibrotic disorders, cartilage (chondral) defects, and osteoarthritis),the modulation of cellular events mediated by Wnt pathway signaling, aswell as genetic diseases and neurological conditions/disorders/diseasesdue to mutations or dysregulation of the Wnt pathway and/or of one ormore of Wnt signaling components. Also provided are methods for treatingWnt-related disease states, as well as neurologicalconditions/disorders/diseases linked to overexpression of DYRK1A.

Background

The Wnt growth factor family includes more than 10 genes identified inthe mouse and at least 19 genes identified in the human. Members of theWnt family of signaling molecules mediate many short- and long-rangepatterning processes during invertebrate and vertebrate development. TheWnt signaling pathway is known for its role in the inductiveinteractions that regulate growth and differentiation, and it also playsroles in the homeostatic maintenance of post-embryonic tissue integrity.Wnt stabilizes cytoplasmic β-catenin, which stimulates the expression ofgenes including c-myc, c jun, fra-1, and cyclin Dl. In addition,misregulation of Wnt signaling can cause developmental defects and isimplicated in the genesis of several human cancers. The Wnt pathway hasalso been implicated in the maintenance of stem or progenitor cells in agrowing list of adult tissues including skin, blood, gut, prostate,muscle, and the nervous system.

Dual specificity tyrosine-phosphorylation-regulated kinase 1A is anenzyme that in humans is encoded by the DYRK1A gene. DYRK1A is a memberof the dual-specificity tyrosine phosphorylation-regulated kinase (DYRK)family. DYRK1A contains a nuclear targeting signal sequence, a proteinkinase domain, a leucine zipper motif, and a highly conservative13-consecutive-histidine repeat. It catalyzes its autophosphorylation onserine/threonine and tyrosine residues. It may play a significant rolein a signaling pathway regulating cell proliferation and may be involvedin brain development. DYRK1A is localized in the Down syndrome criticalregion of chromosome 21, and is considered to be a candidate gene forlearning defects associated with Down syndrome. DYRK1A is also expressedin adult brain neurons, indicating that DYRK1A may play a role in themature central nervous system. Thus, several lines of evidence point tosome synaptic functions of DYRK1A. For instance, it has been found thatDYRK1A phosphorylates and modulates the interaction of severalcomponents of the endocytic protein complex machinery (Dynamin 1,Amphiphysin, and Synaptojanin), suggesting a role in synaptic vesiclerecycling. In addition, a polymorphism (SNP) in DYRK1A was found to beassociated with HIV-1 replication in monocyte-derived macrophages, aswell as with progression to AIDS in two independent cohorts ofHIV-1-infected individuals.

SUMMARY

The present disclosure provides methods and reagents, involvingcontacting a cell with an agent, such as an isoquinoline compound, in asufficient amount to antagonize a Wnt activity, e.g., to reverse orcontrol an aberrant growth state or correct a genetic disorder due tomutations in Wnt signaling components.

The present disclosure also provides methods and reagents, involvingcontacting a cell with an agent, such as an isoquinoline compound, in asufficient amount to antagonize DYRK1A activity, e.g., i) to normalizeprenatal and early postnatal brain development; ii) to improve cognitivefunction in youth and adulthood; and/or iii) to attenuateAlzheimer's-type neurodegeneration.

Some embodiments disclosed herein include Wnt and/or DYRK1A inhibitorscontaining an isoquinoline core. Other embodiments disclosed hereininclude pharmaceutical compositions and methods of treatment using thesecompounds.

One embodiment disclosed herein includes a compound having the structureof Formula I:

as well as prodrugs and pharmaceutically acceptable salts thereof.

In some embodiments of Formula (I):

R¹, R², R⁴, and R⁵ are independently selected from the group consistingof H, halide, unsubstituted —(C₁₋₃ haloalkyl), and unsubstituted —(C₁₋₃alkyl);

R³ is a 5-membered heteroaryl optionally substituted with 1-4 R³⁶;

R⁶ is selected from the group consisting of —(CH₂)(5-memberedheteroaryl) optionally substituted with 1-4 R³⁷ and 5-memberedheteroaryl optionally substituted with 1-4 R³⁸;

with the proviso that R⁶ is not

each R³⁶ is independently selected from the group consisting of H,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R⁴⁴, and-carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein each —(C₁₋₄alkylene) is, independently, optionally substituted with one or moresubstituents as defined anywhere herein;

alternatively, two adjacent R³⁶ taken together form a ring which isselected from the group consisting of -heterocyclyl optionallysubstituted with 1-10 R⁴⁶ and -carbocyclyl optionally substituted with1-12 R⁴⁷;

each R³⁷ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R³⁹, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴⁰;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R³⁸ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R³⁹, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴⁰;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R³⁹ is independently selected from the group consisting ofunsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl),—C(═O)O(R⁴¹), —N(R⁴²)₂, —(C₁₋₄ alkylene)_(p)carbocyclyl optionallysubstituted with 1-12 R⁴³; wherein —(C₁₋₄ alkylene) is, optionallysubstituted with one or more substituents as defined anywhere herein;

each R⁴⁰ is independently selected from the group consisting of halide,unsubstituted —(C₁₋alkyl), unsubstituted —(C₂₋₅ alkenyl), unsubstituted—(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), and —CN;

R⁴¹ is selected from the group consisting of unsubstituted —(C₁₋₉alkyl), unsubstituted —(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl),and unsubstituted —(C₁₋₉ haloalkyl);

each R⁴² is attached to the nitrogen and is selected from the groupconsisting of H, unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉alkenyl), unsubstituted —(C₂₋₉ alkynyl), and unsubstituted —(C₁₋₉haloalkyl);

each R⁴³ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅alkyl), unsubstituted —(C₂₋₅ alkenyl), unsubstituted—(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), and —CN;

each R⁴⁴ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅alkyl), unsubstituted —(C₂₋₅ alkenyl), unsubstituted—(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), —CN, and —(C₁₋₄alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴³; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

each R⁴⁵ independently is selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), —CN, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴³;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R⁴⁶ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), —CN, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴³;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R⁴⁷ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), and —CN;and

each p is independently 0 or 1.

In another embodiment of Formula (I):

R¹, R², R⁴, and R⁵ are independently selected from the group consistingof H, halide, unsubstituted —(C₁₋₃ haloalkyl), and unsubstituted —(C₁₋₃alkyl);

R³ is selected from the group consisting of:

wherein each of R⁷-R³⁵ is, independently, a substituent or a single bondconnecting R³ to the isoquinoline ring; wherein only one of R⁷-R¹⁰ (whenpresent) is a bond, only one of R¹¹-R¹⁴ (when present) is a bond, onlyone of R¹⁵-R¹⁷ (when present) is a bond, only one of R¹⁸-R²⁰ (whenpresent) is a bond, only one of R²¹-R²³ (when present) is a bond, onlyone of R²⁴-R²⁶ (when present) is a bond, only one of R²⁷-R²⁹ (whenpresent) is a bond, only one of R³⁰-R³¹ (when present) is a bond, onlyone of R³²-R³³ (when present) is a bond, and only one of R³⁴-R³⁵ (whenpresent) is a bond; for purposes of clarification, any one of thenitrogen atoms attached to R⁷, R¹¹, R¹⁵, R¹⁸, or R²¹ can serve as thepoint of attachment of R³ to the isoquinoline ring; likewise, any one ofthe carbon atoms attached to R⁸, R⁹, R¹⁰, R¹², R¹³, R¹⁴, R¹⁶, R¹⁷, R¹⁹,R²⁰, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, orR³⁵ can serve as the point of attachment of R³ to the isoquinoline ring;so that:

when the nitrogen atom to which R⁷ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R⁷ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R⁸ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R⁸ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R⁹ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R⁹ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R⁰ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R⁰ is a single bondconnecting R³ to the isoquinoline ring;

when the nitrogen atom to which R¹¹ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R¹¹ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R¹² is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R¹² is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R¹³ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R³ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R¹⁴ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R¹⁴ is a single bondconnecting R³ to the isoquinoline ring;

when the nitrogen atom to which R¹⁵ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R¹⁵ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R¹⁶ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R¹⁶ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R¹⁷ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R¹⁷ is a single bondconnecting R³ to the isoquinoline ring;

when the nitrogen atom to which R¹⁸ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R¹⁸ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R¹⁹ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R¹⁹ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²⁰ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²⁰ is a single bondconnecting R³ to the isoquinoline ring;

when the nitrogen atom to which R²¹ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²¹ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²² is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²² is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²³ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²³ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²⁴ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²⁴ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²⁵ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²⁵ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²⁶ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²⁶ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²⁷ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²⁷ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²⁸ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²⁸ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R²⁹ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R²⁹ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R³⁰ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R³⁰ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R³¹ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R³¹ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R³² is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R³² is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R³³ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R³³ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R³⁴ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R³⁴ is a single bondconnecting R³ to the isoquinoline ring;

when the carbon atom to which R³⁵ is attached serves as the point ofattachment of R³ to the isoquinoline ring, then R³⁵ is a single bondconnecting R³ to the isoquinoline ring;

R⁶ is selected from the group consisting of —(CH₂)(5-memberedheteroaryl) optionally substituted with 1-4 R³⁷, 5-membered heteroaryloptionally substituted with 1-4 R³⁸;

with the proviso that R⁶ is not

R⁷ is selected from the group consisting of a single bond, H,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R⁴⁴, and-carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein —(C₁₋₄alkylene) is, optionally substituted with one or more substituents asdefined anywhere herein;

R⁸, R⁹, and R⁰ are independently selected from the group consisting of asingle bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, one of R⁷ and R⁸, R⁸ and R⁹, or R⁹ and R⁰ are takentogether to form a ring which is selected from the group consisting of-heterocyclyl optionally substituted with 1-10 R⁴⁶ and -carbocyclyloptionally substituted with 1-12 R⁴⁷;

R¹¹ is selected from the group consisting of a single bond, H,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R⁴⁴, and-carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein —(C₁₋₄alkylene) is, optionally substituted with one or more substituents asdefined anywhere herein;

R¹², R¹³, and R¹⁴ are independently selected from the group consistingof a single bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, one of R¹¹ and R¹², R¹² and R¹³, or R¹⁴ and R¹¹ are takentogether to form a ring which is selected from the group consisting of-heterocyclyl optionally substituted with 1-10 R⁴⁶ and -carbocyclyloptionally substituted with 1-12 R⁴⁷;

R¹⁵ is selected from the group consisting of a single bond, H,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R⁴⁴, and-carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein —(C₁₋₄alkylene) is, optionally substituted with one or more substituents asdefined anywhere herein;

R¹⁶ and R¹⁷ are independently selected from the group consisting of asingle bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, one of R¹⁵ and R¹⁶ or R¹⁶ and R¹⁷ are taken together toform a ring which is selected from the group consisting of -heterocyclyloptionally substituted with 1-10 R⁴⁶ and -carbocyclyl optionallysubstituted with 1-12 R⁴⁷;

R¹⁸ is selected from the group consisting of a single bond, H,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R⁴⁴, and-carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein —(C₁₋₄alkylene) is, optionally substituted with one or more substituents asdefined anywhere herein;

R¹⁹ and R²⁰ are independently selected from the group consisting of asingle bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, one of R¹⁸ and R¹⁹ or R¹⁸ and R²⁰ are taken together toform a heterocyclyl optionally substituted with 1-10 R⁴⁶;

R²¹ is selected from the group consisting of a single bond, H,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R⁴⁴, and-carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein —(C₁₋₄alkylene) is, optionally substituted with one or more substituents asdefined anywhere herein;

R²² and R²³ are independently selected from the group consisting of asingle bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, R²² and R²³ are taken together to form a ring which isselected from the group consisting of -heterocyclyl optionallysubstituted with 1-10 R⁴⁶ and -carbocyclyl optionally substituted with1-12 R⁴⁷;

R²⁴, R²⁵, and R²⁶ are independently selected from the group consistingof a single bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, one of R²⁴ and R²⁵ or R²⁵ and R²⁶ are taken together toform a ring which is selected from the group consisting of -heterocyclyloptionally substituted with 1-10 R⁴⁶ and -carbocyclyl optionallysubstituted with 1-12 R⁴⁷;

R²⁷, R²⁸, and R²⁹ are independently selected from the group consistingof a single bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, R²⁷ and R²⁸ are taken together to form a ring which isselected from the group consisting of -heterocyclyl optionallysubstituted with 1-10 R⁴⁶ and -carbocyclyl optionally substituted with1-12 R⁴⁷;

R³⁰ and R³¹ are independently selected from the group consisting of asingle bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, R³⁰ and R³¹ are taken together to form a ring which isselected from the group consisting of -heterocyclyl optionallysubstituted with 1-10 R⁴⁶ and -carbocyclyl optionally substituted with1-12 R⁴⁷;

R³² and R³³ are independently selected from the group consisting of asingle bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

R³⁴ and R³⁵ are independently selected from the group consisting of asingle bond, H, halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted—(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein;

alternatively, R³⁴ and R³⁵ are taken together to form a ring which isselected from the group consisting of -heterocyclyl optionallysubstituted with 1-10 R⁴⁶ and -carbocyclyl optionally substituted with1-12 R⁴⁷;

each R³⁶ is independently selected from the group consisting of H,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R⁴⁴, and-carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein each —(C₁₋₄alkylene) is, independently, optionally substituted with one or moresubstituents as defined anywhere herein;

alternatively, two adjacent R³⁶ taken together form a ring which isselected from the group consisting of -heterocyclyl optionallysubstituted with 1-10 R⁴⁶ and -carbocyclyl optionally substituted with1-12 R⁴⁷;

each R³⁷ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R³⁹, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴⁰;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R³⁸ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄alkylene)_(p)heterocyclyl optionally substituted with 1-10 R³⁹, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴⁰;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R³⁹ is independently selected from the group consisting ofunsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl),—C(═O)O(R⁴¹), —N(R⁴²)₂, —(C₁₋₄ alkylene)_(p)carbocyclyl optionallysubstituted with 1-12 R⁴³; wherein —(C₁₋₄ alkylene) is, optionallysubstituted with one or more substituents as defined anywhere herein;

each R⁴⁰ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), and —CN;

R⁴¹ is selected from the group consisting of unsubstituted —(C₁₋₉alkyl), unsubstituted —(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl),and unsubstituted —(C₁₋₉ haloalkyl); each R⁴² is attached to thenitrogen and is selected from the group consisting of H, unsubstituted—(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉alkynyl), and unsubstituted —(C₁₋₉ haloalkyl);

each R⁴³ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), and —CN;

each R⁴⁴ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), —CN, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴³;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R⁴⁵ independently is selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), —CN, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴³;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R⁴⁶ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), —CN, and—(C₁₋₄ alkylene)_(p)carbocyclyl optionally substituted with 1-12 R⁴³;wherein each —(C₁₋₄ alkylene) is, independently, optionally substitutedwith one or more substituents as defined anywhere herein;

each R⁴⁷ is independently selected from the group consisting of halide,unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅ alkenyl),unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅ haloalkyl), and —CN;

each X is O or S; and

each p is independently 0 or 1.

Some embodiments include stereoisomers and pharmaceutically acceptablesalts of a compound of Formula (I). Some embodiments includepharmaceutically acceptable salts of a compound of Formula (I).

Some embodiments include pro-drugs of a compound of Formula (I).

Some embodiments of the present disclosure include pharmaceuticalcompositions comprising a compound of Formula (I) and a pharmaceuticallyacceptable carrier, diluent, or excipient.

Other embodiments disclosed herein include methods of inhibiting one ormore members of the Wnt pathway, including one or more Wnt proteins byadministering to a patient affected by a disorder or disease in whichaberrant Wnt signaling is implicated, such as cancer and other diseasesassociated with abnormal angiogenesis, cellular proliferation, cellcycling and mutations in Wnt signaling components, a compound accordingto Formula (I). Accordingly, the compounds and compositions providedherein can be used to treat cancer, to reduce or inhibit angiogenesis,to reduce or inhibit cellular proliferation and correct a geneticdisorder due to mutations in Wnt signaling components.

Other embodiments disclosed herein include methods of inhibiting DYRK1Aby administering to a patient affected by a disorder or disease in whichDYRK1A overexpression is implicated, such as Alzheimer's Disease,Amyotrophic Lateral Sclerosis, Down Syndrome, Frontotemporal Dementiawith Parkinsonism-17 (FTDP-17), Lewy body dementia, Parkinson's Disease,Pick's Disease, and additional diseases with pronouncedneurodegeneration such as Autism, Dementia, Epilepsy, Huntington'sDisease, Multiple Sclerosis; diseases and disorders associated withacquired brain injury such as Chronic Traumatic Encephalopathy,Traumatic Brain Injury, Tumor and Stroke.

Non-limiting examples of diseases which can be treated with thecompounds and compositions provided herein include a variety of cancers,diabetic retinopathy, pulmonary fibrosis, rheumatoid arthritis, sepsis,ankylosing spondylitis, psoriasis, scleroderma, mycotic and viralinfections, osteochondrodysplasia, Alzheimer's disease, lung disease,bone/osteoporotic (wrist, spine, shoulder and hip) fractures, articularcartilage (chondral) defects, degenerative disc disease (orintervertebral disc degeneration), polyposis coli,osteoporosis-pseudoglioma syndrome, familial exudativevitreoretinopathy, retinal angiogenesis, early coronary disease,tetra-amelia syndrome, Müllerian-duct regression and virilization,SERKAL syndrome, diabetes mellitus type 2, Fuhrmann syndrome,Al-Awadi/Raas-Rothschild/Schinzel phocomelia syndrome,odonto-onycho-dermal dysplasia, obesity, split-hand/foot malformation,caudal duplication syndrome, tooth agenesis, Wilms tumor, skeletaldysplasia, focal dermal hypoplasia, autosomal recessive anonychia,neural tube defects, alpha-thalassemia (ATRX) syndrome, fragile Xsyndrome, ICF syndrome, Angelman syndrome, Prader-Willi syndrome,Beckwith-Wiedemann Syndrome, Norrie disease, and Rett syndrome.

Some embodiments of the present disclosure include methods to preparecompounds of Formula (I).

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory onlyand are not restrictive of the disclosure, as claimed.

DETAILED DESCRIPTION

Provided herein are compositions and methods for inhibiting one or moremembers of the Wnt pathway, including one or more Wnt proteins. OtherWnt inhibitors and methods for using the same are disclosed in U.S.application Ser. Nos. 13/614,296; 14/019,229; and Ser. No. 14/664,517,all of which are incorporated by reference in their entirety herein.

Provided herein are compositions and methods for inhibiting DYRK1A.Other DYRK1A inhibitors and methods for using the same are disclosed inU.S. application Ser. No. 14/664,517, which is incorporated by referencein its entirety herein.

Some embodiments provided herein relate to a method for treating adisease including, but not limited to, neurological diseases ordisorders, cancers, chronic inflammation, diabetic retinopathy,pulmonary fibrosis, rheumatoid arthritis, sepsis, ankylosingspondylitis, psoriasis, scleroderma, mycotic and viral infections, boneand cartilage diseases, lung disease, osteoarthritis, articularcartilage (chondral) defects, degenerative disc disease (orintervertebral disc degeneration), polyposis coli, bone density andvascular defects in the eye (Osteoporosis-pseudoglioma Syndrome, OPPG),familial exudative vitreoretinopathy, retinal angiogenesis, earlycoronary disease, tetra-amelia, Müllerian-duct regression andvirilization, SERKAL syndrome, type II diabetes, Fuhrmann syndrome,Al-Awadi/Raas-Rothschild/Schinzel phocomelia syndrome,odonto-onycho-dermal dysplasia, obesity, split-hand/foot malformation,caudal duplication, tooth agenesis, Wilms tumor, skeletal dysplasia,focal dermal hypoplasia, autosomal recessive anonychia, neural tubedefects, alpha-thalassemia (ATRX) syndrome, fragile X syndrome, ICFsyndrome, Angelman's syndrome, Prader-Willi syndrome, Beckwith-WiedemannSyndrome, Norrie disease, and Rett syndrome.

In some embodiments, non-limiting examples of bone and cartilagediseases which can be treated with the compounds and compositionsprovided herein include bone spur (osteophytes), craniosynostosis,fibrodysplasia ossificans progressive, fibrous dysplasia, giant celltumor of bone, hip labral tear, meniscal tears, osteoarthritis,articular cartilage (chondral) defects, degenerative disc disease (orintervertebral disc degeneration), osteochondritis dissecans,osteochondroma (bone tumor), osteopetrosis, relapsing polychondritis,and Salter-Harris fractures.

In some embodiments, non-limiting examples of a neurological disease ordisorder associated with tau protein, amyloid or alpha-synucleinpathology which can be treated with the compounds and compositionsprovided herein include, but are not limited to, Alzheimer's Disease,Amyotrophic Lateral Sclerosis, Down Syndrome, Frontotemporal Dementiawith Parkinsonism-17 (FTDP-17), Lewy body dementia, Parkinson's Disease,Pick's Disease, and additional diseases with pronouncedneurodegeneration such as Autism, Dementia, Epilepsy, Huntington'sDisease, Multiple Sclerosis; diseases and disorders associated withacquired brain injury such as Chronic Traumatic Encephalopathy,Traumatic Brain Injury, Tumor, and Stroke.

In some embodiments, non-limiting examples of diseases in which chronicinflammation is involved which can be treated with the compounds andcompositions provided herein include eye disorders, joint pain,arthritis (rheumatoid, osteo, psoriatic gout), cancers (colon, breast,lung, pancreas, and others), gastrointestinal disorders (ulcerativecolitis and inflammatory bowel diseases), pulmonary disorders (chronicobstructive pulmonary disorder and asthma), allergies, skin disorders(atopic dermatitis and psoriasis), diabetes, pancreatitis, tendonitis,hepatitis, heart disease, myocarditis, stroke, lupus, and neurologicaldisorders such as multiple sclerosis, Parkinson's and dementia includingAlzheimer's disease.

In some embodiments, non-limiting examples of cancers which can betreated with the compounds and compositions provided herein includecolon, ovarian, pancreatic, breast, liver, prostate, and hematologiccancers.

In some embodiments, pharmaceutical compositions are provided that areeffective for treatment of a disease of an animal, e.g., a mammal,caused by either the pathological activation or mutations of the Wntpathway or DYRK1A overexpression. The composition includes apharmaceutically acceptable carrier and a compound as described herein.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of ordinary skillin the art to which this disclosure belongs. All patents, applications,published applications, and other publications are incorporated byreference in their entirety. In the event that there is a plurality ofdefinitions for a term herein, those in this section prevail unlessstated otherwise.

As used herein, “alkyl” means a branched, or straight chain chemicalgroup containing only carbon and hydrogen, such as methyl, ethyl,n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl,n-pentyl, iso-pentyl, sec-pentyl and neo-pentyl. Alkyl groups can eitherbe unsubstituted or substituted with one or more substituents. In someembodiments, alkyl groups include 1 to 9 carbon atoms (for example, 1 to6 carbon atoms, 1 to 4 carbon atoms, or 1 to 2 carbon atoms).

As used herein, “alkenyl” means a straight or branched chain chemicalgroup containing only carbon and hydrogen and containing at least onecarbon-carbon double bond, such as ethenyl, 1-propenyl, 2-propenyl,2-methyl-1-propenyl, 1-butenyl, 2-butenyl, and the like. In variousembodiments, alkenyl groups can either be unsubstituted or substitutedwith one or more substituents. Typically, alkenyl groups will comprise 2to 9 carbon atoms (for example, 2 to 6 carbon atoms, 2 to 4 carbonatoms, or 2 carbon atoms).

As used herein, “alkynyl” means a straight or branched chain chemicalgroup containing only carbon and hydrogen and containing at least onecarbon-carbon triple bond, such as ethynyl, 1-propynyl, 1-butynyl,2-butynyl, and the like. In various embodiments, alkynyl groups caneither be unsubstituted or substituted with one or more substituents.Typically, alkynyl groups will comprise 2 to 9 carbon atoms (forexample, 2 to 6 carbon atoms, 2 to 4 carbon atoms, or 2 carbon atoms).

As used herein, “alkylene” means a bivalent branched, or straight chainchemical group containing only carbon and hydrogen, such as methylene,ethylene, n-propylene, iso-propylene, n-butylene, iso-butylene,sec-butylene, tert-butylene, n-pentylene, iso-pentylene, sec-pentyleneand neo-pentylene. Alkylene groups can either be unsubstituted orsubstituted with one or more substituents. In some embodiments, alkylenegroups include 1 to 9 carbon atoms (for example, 1 to 6 carbon atoms, 1to 4 carbon atoms, or 1 to 2 carbon atoms).

As used herein, “alkenylene” means a bivalent branched, or straightchain chemical group containing only carbon and hydrogen and containingat least one carbon-carbon double bond, such as ethenylene,1-propenylene, 2-propenylene, 2-methyl-1-propenylene, 1-butenylene,2-butenylene, and the like. In various embodiments, alkenylene groupscan either be unsubstituted or substituted with one or moresubstituents. Typically, alkenylene groups will comprise 2 to 9 carbonatoms (for example, 2 to 6 carbon atoms, 2 to 4 carbon atoms, or 2carbon atoms).

As used herein, “alkynylene” means a bivalent branched, or straightchain chemical group containing only carbon and hydrogen and containingat least one carbon-carbon triple bond, such as ethynylene,1-propynylene, 1-butynylene, 2-butynylene, and the like. In variousembodiments, alkynylene groups can either be unsubstituted orsubstituted with one or more substituents. Typically, alkynylene groupswill comprise 2 to 9 carbon atoms (for example, 2 to 6 carbon atoms, 2to 4 carbon atoms, or 2 carbon atoms).

As used herein, “alkoxy” means an alkyl-O— group in which the alkylgroup is as described herein. Exemplary alkoxy groups include methoxy,ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy, pentoxy,hexoxy and heptoxy, and also the linear or branched positional isomersthereof.

As used herein, “haloalkoxy” means a haloalkyl-O— group in which thehaloalkyl group is as described herein. Exemplary haloalkoxy groupsinclude fluoromethoxy, difluoromethoxy, trifluoromethoxy, and also thelinear or branched positional isomers thereof.

As used herein, “carbocyclyl” means a cyclic ring system containing onlycarbon atoms in the ring system backbone, such as cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, and cyclohexenyl. Carbocyclyls mayinclude multiple fused rings. Carbocyclyls may have any degree ofsaturation provided that none of the rings in the ring system arearomatic. Carbocyclyl groups can either be unsubstituted or substitutedwith one or more substituents. In some embodiments, carbocyclyl groupsinclude 3 to 10 carbon atoms, for example, 3 to 6 carbon atoms.

As used herein, “aryl” means a mono-, bi-, tri- or polycyclic group withonly carbon atoms present in the ring backbone having 5 to 14 ringatoms, alternatively 5, 6, 9, or 10 ring atoms; and having 6, 10, or 14pi electrons shared in a cyclic array; wherein at least one ring in thesystem is aromatic. Aryl groups can either be unsubstituted orsubstituted with one or more substituents. Examples of aryl includephenyl, naphthyl, tetrahydronaphthyl, 2,3-dihydro-1H-indenyl, andothers. In some embodiments, the aryl is phenyl.

As used herein, “arylalkylene” means an aryl-alkylene- group in whichthe aryl and alkylene moieties are as previously described. In someembodiments, arylalkylene groups contain a C₁₋₄alkylene moiety.Exemplary arylalkylene groups include benzyl and 2-phenethyl.

As used herein, the term “heteroaryl” means a mono-, bi-, tri- orpolycyclic group having 5 to 14 ring atoms, alternatively 5, 6, 9, or 10ring atoms; and having 6, 10, or 14 pi electrons shared in a cyclicarray; wherein at least one ring in the system is aromatic, and at leastone ring in the system contains one or more heteroatoms independentlyselected from the group consisting of N, O, and S. Heteroaryl groups caneither be unsubstituted or substituted with one or more substituents.Examples of heteroaryl include thienyl, pyridinyl, furyl, oxazolyl,oxadiazolyl, pyrrolyl, imidazolyl, triazolyl, thiodiazolyl, pyrazolyl,isoxazolyl, thiadiazolyl, pyranyl, pyrazinyl, pyrimidinyl, pyridazinyl,triazinyl, thiazolyl benzothienyl, benzoxadiazolyl, benzofuranyl,benzimidazolyl, benzotriazolyl, cinnolinyl, indazolyl, indolyl,isoquinolinyl, isothiazolyl, naphthyridinyl, purinyl, thienopyridinyl,pyrido[2,3-d]pyrimidinyl, pyrrolo[2,3-b]pyridinyl, quinazolinyl,quinolinyl, thieno[2,3-c]pyridinyl, pyrazolo[3,4-b]pyridinyl,pyrazolo[3,4-c]pyridinyl, pyrazolo[4,3-c]pyridine,pyrazolo[4,3-b]pyridinyl, tetrazolyl, chromane,2,3-dihydrobenzo[b][1,4]dioxine, benzo[d][1,3]dioxole,2,3-dihydrobenzofuran, tetrahydroquinoline,2,3-dihydrobenzo[b][1,4]oxathiine, isoindoline, and others. In someembodiments, the heteroaryl is selected from thienyl, pyridinyl, furyl,pyrazolyl, imidazolyl, isoindolinyl, pyranyl, pyrazinyl, andpyrimidinyl.

As used herein, “halo”, “halide” or “halogen” is a chloro, bromo,fluoro, or iodo atom radical. In some embodiments, a halo is a chloro,bromo or fluoro. For example, a halide can be fluoro.

As used herein, “haloalkyl” means a hydrocarbon substituent, which is alinear or branched, alkyl, alkenyl or alkynyl substituted with one ormore chloro, bromo, fluoro, and/or iodo atom(s). In some embodiments, ahaloalkyl is a fluoroalkyls, wherein one or more of the hydrogen atomshave been substituted by fluoro. In some embodiments, haloalkyls are of1 to about 3 carbons in length (e.g., 1 to about 2 carbons in length or1 carbon in length). The term “haloalkylene” means a diradical variantof haloalkyl, and such diradicals may act as spacers between radicals,other atoms, or between a ring and another functional group.

As used herein, “heterocyclyl” means a nonaromatic cyclic ring systemcomprising at least one heteroatom in the ring system backbone.Heterocyclyls may include multiple fused rings. Heterocyclyls may besubstituted or unsubstituted with one or more substituents. In someembodiments, heterocycles have 3-11 members. In six membered monocyclicheterocycles, the heteroatom(s) are selected from one to three of O, Nor S, and wherein when the heterocycle is five membered, it can have oneor two heteroatoms selected from O, N, or S. Examples of heterocyclylinclude azirinyl, aziridinyl, azetidinyl, oxetanyl, thietanyl,1,4,2-dithiazolyl, dihydropyridinyl, 1,3-dioxanyl, 1,4-dioxanyl,1,3-dioxolanyl, morpholinyl, thiomorpholinyl, piperazinyl, pyranyl,pyrrolidinyl, tetrahydrofuryl, tetrahydropyridinyl, oxazinyl, thiazinyl,thiinyl, thiazolidinyl, isothiazolidinyl, oxazolidinyl, isoxazolidinyl,piperidinyl, pyrazolidinyl imidazolidinyl, thiomorpholinyl, and others.In some embodiments, the heterocyclyl is selected from azetidinyl,morpholinyl, piperazinyl, pyrrolidinyl, and tetrahydropyridinyl.

As used herein, “monocyclic heterocyclyl” means a single nonaromaticcyclic ring comprising at least one heteroatom in the ring systembackbone. Heterocyclyls may be substituted or unsubstituted with one ormore substituents. In some embodiments, heterocycles have 3-7 members.In six membered monocyclic heterocycles, the heteroatom(s) are selectedfrom one to three of O, N or S, and wherein when the heterocycle is fivemembered, it can have one or two heteroatoms selected from O, N, or S.Examples of heterocyclyls include azirinyl, aziridinyl, azetidinyl,oxetanyl, thietanyl, 1,4,2-dithiazolyl, dihydropyridinyl, 1,3-dioxanyl,1,4-dioxanyl, 1,3-dioxolanyl, morpholinyl, thiomorpholinyl, piperazinyl,pyranyl, pyrrolidinyl, tetrahydrofuryl, tetrahydropyridinyl, oxazinyl,thiazinyl, thiinyl, thiazolidinyl, isothiazolidinyl, oxazolidinyl,isoxazolidinyl, piperidinyl, pyrazolidinyl imidazolidinyl,thiomorpholinyl, and others.

As used herein, “bicyclic heterocyclyl” means a nonaromatic bicyclicring system comprising at least one heteroatom in the ring systembackbone. Bicyclic heterocyclyls may be substituted or unsubstitutedwith one or more substituents. In some embodiments, bicyclicheterocycles have 4-11 members with the heteroatom(s) being selectedfrom one to five of O, N or S. Examples of bicyclic heterocyclylsinclude 2-azabicyclo[1.1.0]butane, 2-azabicyclo[2.1.0]pentane,2-azabicyclo[1.1.1]pentane, 3-azabicyclo[3.1.0]hexane,5-azabicyclo[2.1.1]hexane, 3-azabicyclo[3.2.0]heptane,octahydrocyclopenta[c]pyrrole, 3-azabicyclo[4.1.0]heptane,7-azabicyclo[2.2.1]heptane, 6-azabicyclo[3.1.1]heptane,7-azabicyclo[4.2.0]octane, 2-azabicyclo[2.2.2]octane, and the like.

As used herein, “spirocyclic heterocyclyl” means a nonaromatic bicyclicring system comprising at least one heteroatom in the ring systembackbone and with the rings connected through just one atom. Spirocyclicheterocyclyls may be substituted or unsubstituted with one or moresubstituents. In some embodiments, spirocyclic heterocycles have 5-11members with the heteroatom(s) being selected from one to five of O, Nor S. Examples of spirocyclic heterocyclyls include2-azaspiro[2.2]pentane, 4-azaspiro[2.5]octane, 1-azaspiro[3.5]nonane,2-azaspiro[3.5]nonane, 7-azaspiro[3.5]nonane, 2-azaspiro[4.4]nonane,6-azaspiro[2.6]nonane, 1,7-diazaspiro[4.5]decane,2,5-diazaspiro[3.6]decane, and the like.

The term “substituted” refers to moieties having substituents replacinga hydrogen on one or more non-hydrogen atoms of the molecule. It will beunderstood that “substitution” or “substituted with” includes theimplicit proviso that such substitution is in accordance with permittedvalence of the substituted atom and the substituent, and that thesubstitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, etc. Substituents can include, for example,—(C₁₋₉ alkyl) optionally substituted with one or more of hydroxyl, —NH₂,—NH(C₁₋₃ alkyl), and —N(C₁₋₃ alkyl)₂; —(C₁₋₉ haloalkyl); a halide; ahydroxyl; a carbonyl [such as —C(O)OR, and —C(O)R]; a thiocarbonyl [suchas —C(S)OR, —C(O)SR, and —C(S)R]; —(C₁₋₉ alkoxy) optionally substitutedwith one or more of halide, hydroxyl, —NH₂, —NH(C₁₋₃ alkyl), and —N(C₁₋₃alkyl)₂; —OPO(OH)₂; a phosphonate [such as —PO(OH)₂ and —PO(OR′)₂];—OPO(OR′)R″; —NRR′; —C(O)NRR′; —C(NR)NR′R″; —C(NR′)R″; a cyano; a nitro;an azido; —SH; —S—R; —OSO₂(OR); a sulfonate [such as —SO₂(OH) and—SO₂(OR)]; —SO₂NR′R″; and —SO₂R; in which each occurrence of R, R′ andR″ are independently selected from H; —(C₁₋₉ alkyl); C₆₋₁₀ aryloptionally substituted with from 1-3R′″; 5-10 membered heteroaryl havingfrom 1-4 heteroatoms independently selected from N, O, and S andoptionally substituted with from 1-3 R′″; C₃₋₇ carbocyclyl optionallysubstituted with from 1-3 R′″; and 3-8 membered heterocyclyl having from1-4 heteroatoms independently selected from N, O, and S and optionallysubstituted with from 1-3 R′″; wherein each R′″ is independentlyselected from —(C₁₋₆ alkyl), —(C₁₋₆ haloalkyl), a halide (e.g., F), ahydroxyl, —C(O)OR, —C(O)R, —(C₁₋₆ alkoxyl), —NRR′, —C(O)NRR′, and acyano, in which each occurrence of R and R′ is independently selectedfrom H and —(C₁₋₆ alkyl). In some embodiments, the substituent isselected from —(C₁₋₆ alkyl), —(C₁₋₆ haloalkyl), a halide (e.g., F), ahydroxyl, —C(O)OR, —C(O)R, —(C₁₋₆ alkoxyl), —NRR′, —C(O)NRR′, and acyano, in which each occurrence of R and R′ is independently selectedfrom H and —(C₁₋₆ alkyl).

As used herein, when two groups are indicated to be “linked” or “bonded”to form a “ring”, it is to be understood that a bond is formed betweenthe two groups and may involve replacement of a hydrogen atom on one orboth groups with the bond, thereby forming a carbocyclyl, heterocyclyl,aryl, or heteroaryl ring. The skilled artisan will recognize that suchrings can and are readily formed by routine chemical reactions. In someembodiments, such rings have from 3-7 members, for example, 5 or 6members.

The skilled artisan will recognize that some chemical structuresdescribed herein may be represented on paper by one or more otherresonance forms; or may exist in one or more other tautomeric forms,even when kinetically, the artisan recognizes that such tautomeric formsrepresent only a very small portion of a sample of such compound(s).Such compounds are clearly contemplated within the scope of thisdisclosure, though such resonance forms or tautomers are not explicitlyrepresented herein.

The compounds provided herein may encompass various stereochemicalforms. The compounds also encompass diastereomers as well as opticalisomers, e.g., mixtures of enantiomers including racemic mixtures, aswell as individual enantiomers and diastereomers, which arise as aconsequence of structural asymmetry in certain compounds. Separation ofthe individual isomers or selective synthesis of the individual isomersis accomplished by application of various methods which are well knownto practitioners in the art. Unless otherwise indicated, when adisclosed compound is named or depicted by a structure withoutspecifying the stereochemistry and has one or more chiral centers, it isunderstood to represent all possible stereoisomers of the compound.

The present disclosure includes all pharmaceutically acceptableisotopically labeled compounds of Formula I wherein one or more atomsare replaced by atoms having the same atomic number, but an atomic massor mass number different from the atomic mass or mass number whichpredominates in nature. Examples of isotopes suitable for inclusion inthe compounds of the disclosure include, but are not limited to,isotopes of hydrogen, such as ²H (deuterium) and ³H (tritium), carbon,such as ¹¹C, ¹³C and ¹⁴C, chlorine, such as ³⁶Cl, fluorine, such as^(1s)F, iodine, such as ¹²³I and ¹²⁵I, nitrogen, such as ¹³N and ¹⁵N,oxygen, such as ¹⁵O, 17O and ¹⁸O, phosphorus, such as ³²P, and sulfur,such as ³⁵S.

The term “administration” or “administering” refers to a method ofproviding a dosage of a compound or pharmaceutical composition to avertebrate or invertebrate, including a mammal, a bird, a fish, or anamphibian, where the method is, e.g., orally, subcutaneously,intravenously, intralymphatic, intranasally, topically, transdermally,intraperitoneally, intramuscularly, intrapulmonarilly, vaginally,rectally, ontologically, neuro-otologically, intraocularly,subconjuctivally, via anterior eye chamber injection, intravitreally,intraperitoneally, intrathecally, intracystically, intrapleurally, viawound irrigation, intrabuccally, intra-abdominally, intra-articularly,intra-aurally, intrabronchially, intracapsularly, intrameningeally, viainhalation, via endotracheal or endobronchial instillation, via directinstillation into pulmonary cavities, intraspinally, intrasynovially,intrathoracically, via thoracostomy irrigation, epidurally,intratympanically, intracisternally, intravascularly,intraventricularly, intraosseously, via irrigation of infected bone, orvia application as part of any admixture with a prosthetic device. Themethod of administration can vary depending on various factors, e.g.,the components of the pharmaceutical composition, the site of thedisease, the disease involved, and the severity of the disease.

A “diagnostic” as used herein is a compound, method, system, or devicethat assists in the identification or characterization of a health ordisease state. The diagnostic can be used in standard assays as is knownin the art.

The term “mammal” is used in its usual biological sense. Thus, itspecifically includes humans, cattle, horses, monkeys, dogs, cats, mice,rats, cows, sheep, pigs, goats, and non-human primates, but alsoincludes many other species.

The term “pharmaceutically acceptable carrier”, “pharmaceuticallyacceptable diluent” or “pharmaceutically acceptable excipient” includesany and all solvents, co-solvents, complexing agents, dispersion media,coatings, isotonic and absorption delaying agents and the like which arenot biologically or otherwise undesirable. The use of such media andagents for pharmaceutically active substances is well known in the art.Except insofar as any conventional media or agent is incompatible withthe active ingredient, its use in the therapeutic compositions iscontemplated. Supplementary active ingredients can also be incorporatedinto the compositions. In addition, various adjuvants such as arecommonly used in the art may be included. These and other such compoundsare described in the literature, e.g., in the Merck Index, Merck &Company, Rahway, N.J. Considerations for the inclusion of variouscomponents in pharmaceutical compositions are described, e.g., inBrunton et al. (Eds.) (2017); Goodman and Gilman's: The PharmacologicalBasis of Therapeutics, 13th Ed., The McGraw-Hill Companies.

The term “pharmaceutically acceptable salt” refers to salts that retainthe biological effectiveness and properties of the compounds providedherein and, which are not biologically or otherwise undesirable. In manycases, the compounds provided herein are capable of forming acid and/orbase salts by virtue of the presence of amino and/or carboxyl groups orgroups similar thereto. Many such salts are known in the art, forexample, as described in WO 87/05297. Pharmaceutically acceptable acidaddition salts can be formed with inorganic acids and organic acids.Inorganic acids from which salts can be derived include, for example,hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. Organic acids from which salts can bederived include, for example, acetic acid, propionic acid, glycolicacid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinicacid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamicacid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceuticallyacceptable base addition salts can be formed with inorganic and organicbases. Inorganic bases from which salts can be derived include, forexample, sodium, potassium, lithium, ammonium, calcium, magnesium, iron,zinc, copper, manganese, aluminum, and the like; particularly preferredare the ammonium, potassium, sodium, calcium, and magnesium salts.Organic bases from which salts can be derived include, for example,primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines, basic ionexchange resins, and the like, specifically such as isopropylamine,trimethylamine, diethylamine, triethylamine, tripropylamine, andethanolamine.

“Patient” as used herein, means a human or a non-human mammal, e.g., adog, a cat, a mouse, a rat, a cow, a sheep, a pig, a goat, a non-humanprimate, or a bird, e.g., a chicken, as well as any other vertebrate orinvertebrate. In some embodiments, the patient is a human.

A “therapeutically effective amount” of a compound as provided herein isone which is sufficient to achieve the desired physiological effect andmay vary according to the nature and severity of the disease condition,and the potency of the compound. “Therapeutically effective amount” isalso intended to include one or more of the compounds of Formula I incombination with one or more other agents that are effective to treatthe diseases and/or conditions described herein. The combination ofcompounds can be a synergistic combination. Synergy, as described, forexample, by Chou and Talalay, Advances in Enzyme Regulation (1984), 22,27-55, occurs when the effect of the compounds when administered incombination is greater than the additive effect of the compounds whenadministered alone as a single agent. In general, a synergistic effectis most clearly demonstrated at sub-optimal concentrations of thecompounds. It will be appreciated that different concentrations may beemployed for prophylaxis than for treatment of an active disease. Thisamount can further depend upon the patient's height, weight, sex, ageand medical history.

A therapeutic effect relieves, to some extent, one or more of thesymptoms of the disease.

“Treat,” “treatment,” or “treating,” as used herein refers toadministering a compound or pharmaceutical composition as providedherein for therapeutic purposes. The term “therapeutic treatment” refersto administering treatment to a patient already suffering from a diseasethus causing a therapeutically beneficial effect, such as amelioratingexisting symptoms, ameliorating the underlying metabolic causes ofsymptoms, postponing or preventing the further development of adisorder, and/or reducing the severity of symptoms that will or areexpected to develop.

“Drug-eluting” and/or controlled release as used herein refers to anyand all mechanisms, e.g., diffusion, migration, permeation, and/ordesorption by which the drug(s) incorporated in the drug-elutingmaterial pass therefrom over time into the surrounding body tissue.

“Drug-eluting material” and/or controlled release material as usedherein refers to any natural, synthetic or semi-synthetic materialcapable of acquiring and retaining a desired shape or configuration andinto which one or more drugs can be incorporated and from whichincorporated drug(s) are capable of eluting over time.

“Elutable drug” as used herein refers to any drug or combination ofdrugs having the ability to pass over time from the drug-elutingmaterial in which it is incorporated into the surrounding areas of thebody.

Compounds

The compounds and compositions described herein can be used asanti-proliferative agents, e.g., anti-cancer and anti-angiogenesisagents, and/or as inhibitors of the Wnt signaling pathway, e.g., fortreating diseases or disorders associated with aberrant Wnt signaling.In addition, the compounds can be used as inhibitors of one or morekinases, kinase receptors, or kinase complexes. Such compounds andcompositions are also useful for controlling cellular proliferation,differentiation, and/or apoptosis.

The compounds and compositions described herein can be used to inhibitDYRK1A for treating a disorder or disease in which DYRK1A overexpressionis implicated, such as Alzheimer's Disease, Amyotrophic LateralSclerosis, Down Syndrome, Frontotemporal Dementia with Parkinsonism-17(FTDP-17), Lewy body dementia, Parkinson's Disease, Pick's Disease, andadditional diseases with pronounced neurodegeneration such as Autism,Dementia, Epilepsy, Huntington's Disease, Multiple Sclerosis; diseasesand disorders associated with acquired brain injury such as ChronicTraumatic Encephalopathy, Traumatic Brain Injury, Tumor, and Stroke.

Some embodiments of the present disclosure include compounds of FormulaI:

or salts, pharmaceutically acceptable salts, or prodrugs thereof.

In some embodiments, R¹, R², R⁴, and R⁵ are independently selected fromthe group consisting of H, halide, unsubstituted —(C₁₋₃ haloalkyl), andunsubstituted —(C₁₋₃ alkyl).

In some embodiments, R¹, R², R⁴, and R⁵ are independently selected fromthe group consisting of H and halide (e.g., F, Cl, Br, I).

In some embodiments, R¹, R², R⁴, and R⁵ are independently selected fromthe group consisting of H and F.

In some embodiments, R¹, R², R⁴, and R⁵ are all H.

In some embodiments, R¹ is F, and R², R⁴, and R⁵ are all H.

In some embodiments, R² is F, and R¹, R⁴, and R⁵ are all H.

In some embodiments, R⁴ is F, and R¹, R², and R⁵ are all H.

In some embodiments, R⁵ is F, and R¹, R², and R⁴ are all H.

In some embodiments, R³ is a 5-membered heteroaryl ring optionallysubstituted as defined anywhere herein.

In some embodiments, R³ is 5-membered heteroaryl ring optionallysubstituted with 1-4 (e.g., 1-3, 1-2, 1) R³⁶;

In some embodiments, R³ is selected from the group consisting of:furanyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1) R³⁶,thiophenyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1) R³⁶,pyrrolyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1) R³⁶,

wherein each m is independently 1 to 4 (e.g., 1-3, 1-2, 1).

In some embodiments, R³ is selected from the group consisting of:

wherein each of R⁷-R³⁵ is, independently, a substituent as definedanywhere herein or a single bond connecting R³ to the isoquinoline ring;wherein only one of R⁷-R¹⁰ (when present) is a bond, only one of R¹¹-R¹⁴(when present) is a bond, only one of R¹⁵-R¹⁷ (when present) is a bond,only one of R¹⁸-R²⁰ (when present) is a bond, only one of R²¹-R²³ (whenpresent) is a bond, only one of R²⁴-R²⁶ (when present) is a bond, onlyone of R²⁷-R²⁹ (when present) is a bond, only one of R³⁰-R³¹ (whenpresent) is a bond, only one of R³²-R³³ (when present) is a bond, andonly one of R³⁴-R³⁵ (when present) is a bond; for purposes ofclarification, any one of the nitrogen atoms attached to R⁷, R¹¹, R¹⁵,R¹⁸, or R²¹ can serve as the point of attachment of R³ to theisoquinoline ring; likewise, any one of the carbon atoms attached to R⁸,R⁹, R¹⁰, R¹², R¹³, R⁴, R¹⁶, R⁷, R¹⁹, R²⁰, R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷,R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, or R³⁵ can serve as the point ofattachment of R³ to the isoquinoline ring.

In some embodiment, —(C₁₋₄ alkylene) is, optionally substituted with oneor more substituents as defined anywhere herein.

In some embodiment, —(C₁₋₄ alkylene) is, optionally substituted with oneor more halides (e.g., F, Cl, Br, I) or one or more unsubstituted —(C₁₋₃alkyl) (e.g., C_(2-3, 1-2, 1) alkyl).

In some embodiment, —(C₁₋₄ alkylene) is, optionally substituted with oneor more halides (e.g., F, Cl, Br, I) and one or more unsubstituted—(C₁₋₃ alkyl) (e.g., C_(2-3, 1-2, 1) alkyl).

In some embodiment, —(C₁₋₄ alkylene) is, optionally substituted with oneor more F or one or more Me.

In some embodiment, —(C₁₋₄ alkylene) is, optionally substituted with oneor more F and one or more Me.

In some embodiment, —(C₁₋₄ alkylene) is —CH₂—.

In some embodiment, —(C₁₋₄ alkylene) is —CH₂CH₂—.

In some embodiment, —(C₁₋₄ alkylene) is —CH₂CH₂CH₂—.

In some embodiment, R⁶ is selected from the group consisting of—(CH₂)(5-membered heteroaryl) optionally substituted with 1-4 R³⁷ and5-membered heteroaryl optionally substituted with 1-4 R³⁸.

In some embodiment, there is the proviso that that R⁶ is not

In some embodiments, R⁶ is selected from the group consisting of:—(CH₂)furanyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1) R³⁷,—(CH₂)thiophenyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1)R³⁷, —(CH₂)pyrrolyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1)R³⁷,

wherein each m is independently 1 to 4 (e.g., 1-3, 1-2, 1).

In some embodiments, R⁶ is selected from the group consisting of:furanyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1) R³⁸,thiophenyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1) R³⁸,pyrrolyl optionally substituted with 1-4 (e.g., 1-3, 1-2, 1) R³⁸,

wherein each m is independently 1 to 4 (e.g., 1-3, 1-2, 1).

In certain embodiments, R³ is selected from the group consisting ofpyrazolyl, imidazolyl, triazolyl, thiadiazolyl, and oxazolyl, eachoptionally substituted with 1-4 R³⁶.

In certain embodiments, R³ is selected from the group consisting ofpyrazol-4-yl, imidazol-5-yl, 1,2,3-triazol-4-yl, thiadiazol-2-yl, andoxazol-5-yl, each optionally substituted with 1-4 R³⁶.

In certain embodiments, R³ is an unsubstituted pyrazol-4-yl.

In certain embodiments, R³ is a pyrazol-4-yl substituted with one —(C₁₋₃alkyl).

In certain embodiments, R³ is a imidazol-5-yl substituted with one—(C₁₋₃ alkyl).

In certain embodiments, R³ is a imidazol-5-yl substituted with two—(C₁₋₃ alkyl).

In certain embodiments, R³ is an unsubstituted 1,2,3-triazol-4-yl.

In certain embodiments, R³ is a 1,2,3-triazol-4-yl substituted with one—(C₁₋₃ alkyl).

In certain embodiments, R³ is an unsubstituted thiadiazol-2-yl.

In certain embodiments, R³ is a thiadiazol-2-yl substituted with one—(C₁₋₃ alkyl).

In certain embodiments, R³ is an unsubstituted oxazol-5-yl.

In certain embodiments, R³ is a oxazol-5-yl substituted with one —(C₁₋₃alkyl).

In certain embodiments, R⁶ is —(CH₂)(5-membered heteroaryl) optionallysubstituted with 1-4 R³⁷.

In certain embodiments, R⁶ is —(CH₂)pyrazolyl optionally substitutedwith one —(C₁₋₃ alkyl).

In certain embodiments, R⁶ is —(CH₂)imidazolyl optionally substitutedwith one —(C₁₋₃ alkyl).

In certain embodiments, R⁶ is 5-membered heteroaryl optionallysubstituted with 1-4 R³⁸.

In certain embodiments, R⁶ is thiadiazolyl optionally substituted withone —(C₁₋₃ alkyl) or one heterocyclyl optionally substituted with oneR³⁹.

In certain embodiments, R⁶ is pyrazolyl optionally substituted with one—(C₁₋₃ alkyl) or one heterocyclyl optionally substituted with one R³⁹

In certain embodiments, R⁶ is oxazolyl optionally substituted with one—(C₁₋₃ alkyl) or one heterocyclyl optionally substituted with one R³⁹.

In some embodiments, R³ is

In certain embodiments, R⁹ is a single bond connecting R³ to theisoquinoline ring, i.e., R has the following formula:

In certain embodiments, R⁹ is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In some embodiments, R³ is

and n is 1 to 3.

In some embodiments, R⁷ is selected from the group consisting of asingle bond, H, unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein—(C₁₋₄ alkylene) is, optionally substituted with one or moresubstituents as defined anywhere herein.

In some embodiments, R⁷ is selected from the group consisting of H,unsubstituted —(C₁₋₃ alkyl), unsubstituted —(C₁₋₂ haloalkyl), and —(C₃₋₄carbocyclyl) optionally substituted with 1-2 R⁴⁵.

In some embodiments, R⁷ is selected from the group consisting of H, Me,—CF₃, and cyclopropyl optionally substituted with 1-2 R⁴⁵.

In some embodiments, R⁷ is selected from the group consisting of asingle bond, H, and unsubstituted —(C₁₋₅ alkyl) (e.g.,C_(2-5, 3-5, 4-5, 2-4, 3-4, 2-3, 1-4, 1-3, 1-2, 1) alkyl).

In some embodiments, R⁷ is selected from the group consisting of H andMe.

In some embodiments, R⁷ is Me.

In some embodiments, R⁷ is —CD₃.

In some embodiments, R⁸, R⁹, and R⁰ are independently selected from thegroup consisting of a single bond, H, halide, unsubstituted —(C₁₋₉alkyl), unsubstituted —(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl),unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyloptionally substituted with 1-10 R⁴⁴, and -carbocyclyl optionallysubstituted with 1-12 R⁴⁵; wherein each —(C₁₋₄ alkylene) is,independently, optionally substituted with one or more substituents asdefined anywhere herein.

In some embodiments, one of R⁷ and R⁸, R⁸ and R⁹, or R⁹ and R⁰ are takentogether to form a ring which is selected from the group consisting of-heterocyclyl optionally substituted with 1-10 R⁴⁶ and -carbocyclyloptionally substituted with 1-12 R⁴⁷.

In some embodiments, R⁸, R⁹, and R¹¹ are independently selected from thegroup consisting of a single bond, H, and unsubstituted —(C_(1-s) alkyl)(e.g., C_(2-5, 3-5, 4-5, 2-4, 3-4, 2-3, 1-4, 1-3, 1-2, 1) alkyl).

In some embodiments, R⁸, R⁹, and R¹⁰ are independently selected from thegroup consisting of a single bond, H, and Me.

In some embodiments, R⁹ is a single bond connecting R³ to theisoquinoline ring and R⁸ and R¹⁰ are independently selected from thegroup consisting of H and Me.

In some embodiments, R⁹ is a single bond and R⁸ and R¹⁰ are both H.

In some embodiments, R³ is

In certain embodiments, R¹² is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In certain embodiments, R¹² is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In some embodiments, R³ is

and n is 1 to 3.

In some embodiments, R¹¹ is selected from the group consisting of asingle bond, H, unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein—(C₁₋₄ alkylene) is, optionally substituted with one or moresubstituents as defined anywhere herein.

In some embodiments, R¹¹ is selected from the group consisting of H,unsubstituted —(C₁₋₃ alkyl), unsubstituted —(C₁₋₂ haloalkyl), and —(C₃₋₄carbocyclyl) optionally substituted with 1-2 R⁴⁵.

In some embodiments, R¹¹ is selected from the group consisting of H,methyl, —CF₃, and cyclopropyl optionally substituted with 1-2 R⁴⁵.

In some embodiments, R¹¹ is selected from the group consisting of asingle bond, H, and unsubstituted —(C₁₋₅ alkyl) (e.g.,C_(2-5, 3-5, 4-5, 2-4, 3-4, 2-3, 1-4, 1-3, 1-2, 1) alkyl).

In some embodiments, R¹¹ is selected from the group consisting of H andMe.

In some embodiments, R¹¹ is H.

In some embodiments, R¹¹ is Me.

In some embodiments, R¹¹ is —CD₃.

In some embodiments, R¹², R¹³, and R¹⁴ are independently selected fromthe group consisting of a single bond, H, halide, unsubstituted —(C₁₋₉alkyl), unsubstituted —(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl),unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyloptionally substituted with 1-10 R⁴⁴, and -carbocyclyl optionallysubstituted with 1-12 R⁴⁵; wherein each —(C₁₋₄ alkylene) is,independently, optionally substituted with one or more substituents asdefined anywhere herein.

In some embodiments, one of R¹¹ and R¹², R¹² and R¹³, or R¹⁴ and R¹¹ aretaken together to form a ring which is selected from the groupconsisting of -heterocyclyl optionally substituted with 1-10 R⁴⁶ and-carbocyclyl optionally substituted with 1-12 R⁴⁷.

In some embodiments, R¹², R¹³, and R¹⁴ are independently selected fromthe group consisting of a single bond, H, and unsubstituted —(C₁₋₅alkyl) (e.g., C_(2-5, 3-5, 4-5, 2-4, 3-4, 2-3, 1-4, 1-3, 1-2, 1) alkyl).

In some embodiments, R¹², R¹³, and R¹⁴ are independently selected fromthe group consisting of a single bond, H, and Me.

In some embodiments, R¹² is a single bond connecting R³ to theisoquinoline ring and R³ and R¹⁴ are independently selected from thegroup consisting of H and Me.

In some embodiments, R¹² is a single bond connecting R³ to theisoquinoline ring and R³ and R¹⁴ are both H.

In some embodiments, R¹² is a single bond connecting R³ to theisoquinoline ring; R³ is H, and R⁴ is Me.

In some embodiments, R³ is

In certain embodiments, R¹⁶ is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In certain embodiments, R¹⁷ is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In certain embodiments, R¹⁷ is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In some embodiments, R¹⁵ is selected from the group consisting of asingle bond, H, unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein—(C₁₋₄ alkylene) is, optionally substituted with one or moresubstituents as defined anywhere herein.

In some embodiments, R¹⁵ is selected from the group consisting of asingle bond, H, and unsubstituted —(C₁₋₅ alkyl) (e.g.,C_(2-5, 3-5, 4-5, 2-4, 3-4, 2-3, 1-4, 1-3, 12, 1) alkyl).

In some embodiments, R¹⁵ is selected from the group consisting of H andMe.

In some embodiments, R¹⁵ is H.

In some embodiments, R¹⁵ is Me.

In some embodiments, R¹⁶ and R¹⁷ are independently selected from thegroup consisting of a single bond, H, halide, unsubstituted —(C₁₋₉alkyl), unsubstituted —(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl),unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyloptionally substituted with 1-10 R⁴⁴, and -carbocyclyl optionallysubstituted with 1-12 R⁴⁵; wherein each —(C₁₋₄ alkylene) is,independently, optionally substituted with one or more substituents asdefined anywhere herein.

In some embodiments, one of R¹⁵ and R¹⁶ or R¹⁶ and R¹⁷ are takentogether to form a ring which is selected from the group consisting of-heterocyclyl optionally substituted with 1-10 R⁴⁶ and -carbocyclyloptionally substituted with 1-12 R⁴⁷.

In some embodiments, R¹⁶ and R¹⁷ are independently selected from thegroup consisting of a single bond, H, and unsubstituted —(C₁₋₅ alkyl)(e.g., C_(2-5, 3-5, 4-5, 2-4, 3-4, 2-3, 1-4, 1-3, 1-2, 1) alkyl).

In some embodiments, R¹⁶ and R¹⁷ are independently selected from thegroup consisting of a single bond, H, and Me.

In some embodiments, R¹⁷ is a single bond connecting R³ to theisoquinoline ring and R¹⁶ is selected from the group consisting of H andMe.

In some embodiments, R¹⁷ is a single bond connecting R³ to theisoquinoline ring and R¹⁶ is H.

In some embodiments, R³ is

In certain embodiments, R²⁰ is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In certain embodiments, R²⁰ is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In some embodiments, R³ is

and n is 1 to 3.

In some embodiments, R¹⁸ is selected from the group consisting of asingle bond, H, unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; wherein—(C₁₋₄ alkylene) is, optionally substituted with one or moresubstituents as defined anywhere herein.

In some embodiments, R¹⁸ is selected from the group consisting of H,unsubstituted —(C₁₋₃ alkyl), unsubstituted —(C₁₋₂ haloalkyl), and —(C₃₋₄carbocyclyl) optionally substituted with 1-2 R⁴⁵.

In some embodiments, R¹⁸ is selected from the group consisting of H,methyl, —CF₃, and cyclopropyl optionally substituted with 1-2 R⁴⁵.

In some embodiments, R¹⁸ is selected from the group consisting of asingle bond, H, and unsubstituted —(C₁₋₅ alkyl) (e.g.,C_(2-5, 3-5, 4-5, 2-4, 3-4, 2-3, 1-4, 1-3, 1-2, 1) alkyl).

In some embodiments, R¹⁸ is selected from the group consisting of H andMe.

In some embodiments, R¹⁸ is H.

In some embodiments, R¹⁸ is Me.

In some embodiments, R¹⁹ and R²⁰ are independently selected from thegroup consisting of a single bond, H, halide, unsubstituted —(C₁₋₉alkyl), unsubstituted —(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl),unsubstituted —(C₁₋₉ haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyloptionally substituted with 1-10 R⁴⁴, and -carbocyclyl optionallysubstituted with 1-12 R⁴⁵; wherein each —(C₁₋₄ alkylene) is,independently, optionally substituted with one or more substituents asdefined anywhere herein.

In some embodiments, one of R¹⁸ and R¹⁹ or R¹⁸ and R²⁰ are takentogether to form a heterocyclyl optionally substituted with 1-10 R⁴⁶.

In some embodiments, R¹⁹ and R²⁰ are independently selected from thegroup consisting of a single bond, H, and unsubstituted —(C₁₋₅ alkyl)(e.g., C_(2-5, 3-5, 4-5, 2-4, 3-4, 2-3, 1-4, 1-3, 1-2, 1) alkyl).

In some embodiments, R¹⁹ and R²⁰ are independently selected from thegroup consisting of a single bond, H, and Me.

In some embodiments, R²⁰ is a single bond connecting R³ to theisoquinoline ring and R¹⁹ is selected from the group consisting of H andMe.

In some embodiments, R²⁰ is a single bond connecting R³ to theisoquinoline ring and R¹⁹ is H.

In some embodiments, R³ is

In some embodiments, R³ is

and X is S.

In some embodiments, R³ is

and X is O.

In certain embodiments, R²⁷ is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In some embodiments, R³ is

In some embodiments, R³ is

In some embodiments, R²⁸ is selected from the group consisting of H andhalide.

In some embodiments, R²⁸ is selected from the group consisting of H andF.

In some embodiments, R²⁹ is selected from the group consisting of H,halide, unsubstituted —(C₁₋₂ alkyl), and unsubstituted —(C₁₋₂haloalkyl).

In some embodiments, R²⁹ is selected from the group consisting of H, F,methyl, and —CF₃.

In some embodiments, R³ is

In some embodiments, R³ is

and X is S.

In some embodiments, R³ is

and X is O.

In certain embodiments, R³³ is a single bond connecting R³ to theisoquinoline ring, i.e., R³ has the following formula:

In some embodiments, R³ is

In some embodiments, R³ is

In some embodiments, R³² is selected from the group consisting of H,halide, unsubstituted —(C₁₋₂ alkyl), unsubstituted —(C₁₋₂ haloalkyl),and —N(R⁵³)₂.

In some embodiments, R³² is selected from the group consisting of H, F,methyl, —CF₃, —NHMe, and —NMe₂.

In some embodiments, R³² is selected from the group consisting of H andmethyl.

In some embodiments, R³² is methyl.

In some embodiments, each R³⁶ is independently selected from the groupconsisting of H, unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R⁴⁴, and -carbocyclyl optionally substituted with 1-12 R⁴⁵; whereineach —(C₁₋₄ alkylene) is, independently, optionally substituted with oneor more substituents as defined anywhere herein.

In some embodiments, two adjacent R³⁶ taken together form a ring whichis selected from the group consisting of -heterocyclyl optionallysubstituted with 1-10 R⁴⁶ and -carbocyclyl optionally substituted with1-12 R⁴⁷.

In some embodiments, each R³⁷ is independently selected from the groupconsisting of halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R³⁹, and —(C₁₋₄ alkylene)_(p)carbocyclyl optionally substitutedwith 1-12 R⁴⁰; wherein each —(C₁₋₄ alkylene) is, independently,optionally substituted with one or more substituents as defined anywhereherein.

In some embodiments, each R³⁷ is independently selected from the groupconsisting of unsubstituted —(C₁₋₃ alkyl), -heterocyclyl optionallysubstituted with 1-2 R³⁹, and -carbocyclyl optionally substituted with1-2 R⁴⁰.

In some embodiments, each R³⁷ is independently selected from the groupconsisting of Me, Et, iPr, iBu, cyclopropyl,

and q is 0 to 2.

In some embodiments, each R³⁷ is independently selected from the groupconsisting of Me, Et, iPr, iBu, cyclopropyl,

q is 0 to 2.

In some embodiments, each R³⁸ is independently selected from the groupconsisting of halide, unsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉alkenyl), unsubstituted —(C₂₋₉ alkynyl), unsubstituted —(C₁₋₉haloalkyl), —(C₁₋₄ alkylene)_(p)heterocyclyl optionally substituted with1-10 R³⁹, and —(C₁₋₄ alkylene)_(p)carbocyclyl optionally substitutedwith 1-12 R⁴⁰; wherein each —(C₁₋₄ alkylene) is, independently,optionally substituted with one or more substituents as defined anywhereherein.

In some embodiments, each R³⁸ is independently selected from the groupconsisting of Me, Et, iPr, iBu, cyclopropyl,

and q is 0 to 2.

In some embodiments, each R³⁸ is independently selected from the groupconsisting of Me, Et, iPr, iBu, cyclopropyl,

q is 0 to 2.

In some embodiments, each R³⁹ is independently selected from the groupconsisting of unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅alkenyl), unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅haloalkyl), —C(═O)O(R⁴¹), —N(R⁴²)₂, —(C₁₋₄ alkylene)_(p)carbocyclyloptionally substituted with 1-12 R⁴³; wherein —(C₁₋₄ alkylene) is,optionally substituted with one or more substituents as defined anywhereherein.

In some embodiments, each R⁴⁰ is independently selected from the groupconsisting of halide, unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅alkenyl), unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅haloalkyl), and —CN.

In some embodiments, R⁴¹ is selected from the group consisting ofunsubstituted —(C₁₋₉ alkyl), unsubstituted —(C₂₋₉ alkenyl),unsubstituted —(C₂₋₉ alkynyl), and unsubstituted —(C₁₋₉ haloalkyl).

In some embodiments, each R⁴² is attached to the nitrogen and isselected from the group consisting of H, unsubstituted —(C₁₋₉ alkyl),unsubstituted —(C₂₋₉ alkenyl), unsubstituted —(C₂₋₉ alkynyl), andunsubstituted —(C₁₋₉ haloalkyl).

In some embodiments, each R⁴³ is independently selected from the groupconsisting of halide, unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅alkenyl), unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅haloalkyl), and —CN.

In some embodiments, each R⁴⁴ is independently selected from the groupconsisting of halide, unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅alkenyl), unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅haloalkyl), —CN, and —(C₁₋₄ alkylene)_(p)carbocyclyl optionallysubstituted with 1-12 R⁴³; wherein-(C₁₋₄ alkylene) is, optionallysubstituted with one or more substituents as defined anywhere herein.

In some embodiments, each R⁴⁵ independently is selected from the groupconsisting of halide, unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅alkenyl), unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅haloalkyl), —CN, and —(C₁₋₄ alkylene)_(p)carbocyclyl optionallysubstituted with 1-12 R⁴³; wherein each —(C₁₋₄ alkylene) is,independently, optionally substituted with one or more substituents asdefined anywhere herein.

In some embodiments, each R⁴⁶ is independently selected from the groupconsisting of halide, unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅alkenyl), unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅haloalkyl), —CN, and —(C₁₋₄ alkylene)_(p)carbocyclyl optionallysubstituted with 1-12 R⁴³; wherein each —(C₁₋₄ alkylene) is,independently, optionally substituted with one or more substituents asdefined anywhere herein.

In some embodiments, each R⁴⁷ is independently selected from the groupconsisting of halide, unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅alkenyl), unsubstituted —(C₂₋₅ alkynyl), unsubstituted —(C₁₋₅haloalkyl), and —CN.

In some embodiments, each p is independently 0 or 1.

In some embodiments, p is 0.

In some embodiments, p is 1.

In some embodiments, each X is O or S.

Illustrative compounds of Formula (I) are shown in Table 1.

TABLE 1 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

Administration and Pharmaceutical Compositions

Some embodiments include pharmaceutical compositions comprising: (a) atherapeutically effective amount of a compound provided herein, or itscorresponding enantiomer, diastereoisomer or tautomer, orpharmaceutically acceptable salt; and (b) a pharmaceutically acceptablecarrier.

The compounds provided herein may also be useful in combination(administered together or sequentially) with other known agents.

Non-limiting examples of diseases which can be treated with acombination of a compound of Formula (I) and other another active agentare colorectal cancer, ovarian cancer, chronic inflammation, diabeticretinopathy, pulmonary fibrosis, and osteoarthritis. For example, acompound of Formula (I) can be combined with one or morechemotherapeutic compounds.

In some embodiments, colorectal cancer can be treated with a combinationof a compound of Formula (I) and one or more of the following drugs:5-Fluorouracil (5-FU), which can be administered with the vitamin-likedrug leucovorin (also called folinic acid); capecitabine (XELODA®),irinotecan (CAMPOSTAR®), oxaliplatin (ELOXATIN®). Examples ofcombinations of these drugs which could be further combined with acompound of Formula (I) are FOLFOX (5-FU, leucovorin, and oxaliplatin),FOLFIRI (5-FU, leucovorin, and irinotecan), FOLFOXIRI (leucovorin, 5-FU,oxaliplatin, and irinotecan) and CapeOx (Capecitabine and oxaliplatin).For rectal cancer, chemo with 5-FU or capecitabine combined withradiation may be given before surgery (neoadjuvant treatment).

In some embodiments, ovarian cancer can be treated with a combination ofa compound of Formula (I) and one or more of the following drugs:Topotecan, Liposomal doxorubicin (DOXIL®), Gemcitabine (GEMZAR®),Cyclophosphamide (CYTOXAN®), Vinorelbine (NAVELBINE®), Ifosfamide(IFEX®), Etoposide (VP-16), Altretamine (HEXALEN®), Capecitabine(XELODA®), Irinotecan (CPT-11, CAMPTOSAR®), Melphalan, Pemetrexed(ALIMTA®) and Albumin bound paclitaxel (nab-paclitaxel, ABRAXANE®).Examples of combinations of these drugs which could be further combinedwith a compound of Formula (I) are TIP (paclitaxel [Taxol], ifosfamide,and cisplatin), VeIP (vinblastine, ifosfamide, and cisplatin) and VIP(etoposide [VP-16], ifosfamide, and cisplatin).

In some embodiments, a compound of Formula (I) can be used to treatcancer in combination with any of the following methods: (a) Hormonetherapy such as aromatase inhibitors, LHRH [luteinizinghormone-releasing hormone] analogs and inhibitors, and others; (b)Ablation or embolization procedures such as radiofrequency ablation(RFA), ethanol (alcohol) ablation, microwave thermotherapy andcryosurgery (cryotherapy); (c) Chemotherapy using alkylating agents suchas cisplatin and carboplatin, oxaliplatin, mechlorethamine,cyclophosphamide, chlorambucil and ifosfamide; (d) Chemotherapy usinganti-metabolites such as azathioprine and mercaptopurine; (e)Chemotherapy using plant alkaloids and terpenoids such as vincaalkaloids (i.e. Vincristine, Vinblastine, Vinorelbine and Vindesine) andtaxanes; (f) Chemotherapy using podophyllotoxin, etoposide, teniposideand docetaxel; (g) Chemotherapy using topoisomerase inhibitors such asirinotecan, topotecan, amsacrine, etoposide, etoposide phosphate, andteniposide; (h) Chemotherapy using cytotoxic antibiotics such asactinomycin, anthracyclines, doxorubicin, daunorubicin, valrubicin,idarubicin, epirubicin, bleomycin, plicamycin and mitomycin; (i)Chemotherapy using tyrosine-kinase inhibitors such as Imatinib mesylate(GLEEVEC®, also known as STI-571), Gefitinib (Iressa, also known asZD1839), Erlotinib (marketed as TARCEVA®), Bortezomib (VELCADE®),tamoxifen, tofacitinib, crizotinib, Bcl-2 inhibitors (e.g. obatoclax inclinical trials, ABT-263, and Gossypol), PARP inhibitors (e.g. Iniparib,Olaparib in clinical trials), PI3K inhibitors (e.g. perifosine in aphase III trial), VEGF Receptor 2 inhibitors (e.g. Apatinib), AN-152,(AEZS-108), Braf inhibitors (e.g. vemurafenib, dabrafenib and LGX818),MEK inhibitors (e.g. trametinib and MEK162), CDK inhibitors, (e.g.PD-0332991), salinomycin and Sorafenib; (j) Chemotherapy usingmonoclonal antibodies such as Rituximab (marketed as MABTHERA® orRITUXAN®), Trastuzumab (Herceptin also known as ErbB2), Cetuximab(marketed as ERBITUX®), and Bevacizumab (marketed as AVASTIN®); and (k)radiation therapy.

In some embodiments, diabetic retinopathy can be treated with acombination of a compound of Formula (I) and one or more of thefollowing natural supplements: Bilberry, Butcher's broom, Ginkgo, Grapeseed extract, and Pycnogenol (Pine bark).

In some embodiments, idiopathic pulmonary fibrosis/pulmonary fibrosiscan be treated with a combination of a compound of Formula (I) and oneor more of the following drugs: pirfenidone (pirfenidone was approvedfor use in 2011 in Europe under the brand name Esbriet®), prednisone,azathioprine, N-acetylcysteine, interferon-γ 1b, bosentan (bosentan iscurrently being studied in patients with IPF, [The American Journal ofRespiratory and Critical Care Medicine (2011), 184(1), 92-9]),Nintedanib (BIBF 1120 and Vargatef), QAX576 [British Journal ofPharmacology (2011), 163(1), 141-172], and anti-inflammatory agents suchas corticosteroids.

In some embodiments, a compound of Formula (I) can be used to treatidiopathic pulmonary fibrosis/pulmonary fibrosis in combination with anyof the following methods: oxygen therapy, pulmonary rehabilitation andsurgery.

In some embodiments, a compound of Formula (I) can be used to treatosteoarthritis in combination with any of the following methods: (a)Nonsteroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen,naproxen, aspirin and acetaminophen; (b) physical therapy; (c)injections of corticosteroid medications; (d) injections of hyaluronicacid derivatives (e.g. Hyalgan, Synvisc); (e) narcotics, like codeine;(f) in combination with braces and/or shoe inserts or any device thatcan immobilize or support your joint to help you keep pressure off it(e.g., splints, braces, shoe inserts or other medical devices); (g)realigning bones (osteotomy); (h) joint replacement (arthroplasty); and(i) in combination with a chronic pain class.

In some embodiments, macular degeneration can be treated with acombination of a compound of Formula (I) and one or more of thefollowing drugs: Bevacizumab (Avastin®), Ranibizumab (Lucentis®),Pegaptanib (Macugen), Aflibercept (Eylea®), verteporfin (Visudyne®) incombination with photodynamic therapy (PDT) or with any of the followingmethods: (a) in combination with laser to destroy abnormal blood vessels(photocoagulation); and (b) in combination with increased vitamin intakeof antioxidant vitamins and zinc.

In some embodiments, retinitis pigmentosa can be treated with acombination of a compound of Formula (I) and one or more of thefollowing drugs: UF-021 (Ocuseva™), vitamin A palmitate and pikachurinor with any of the following methods: (a) with the Argus® II retinalimplant; and (b) with stem cell and/or gene therapy.

Administration of the compounds disclosed herein or the pharmaceuticallyacceptable salts thereof can be via any of the accepted modes ofadministration, including, but not limited to, orally, subcutaneously,intravenously, intranasally, topically, transdermally,intraperitoneally, intramuscularly, intrapulmonarilly, vaginally,rectally, ontologically, neuro-otologically, intraocularly,subconjuctivally, via anterior eye chamber injection, intravitreally,intraperitoneally, intrathecally, intracystically, intrapleurally, viawound irrigation, intrabuccally, intra-abdominally, intra-articularly,intra-aurally, intrabronchially, intracapsularly, intrameningeally, viainhalation, via endotracheal or endobronchial instillation, via directinstillation into pulmonary cavities, intraspinally, intrasynovially,intrathoracically, via thoracostomy irrigation, epidurally,intratympanically, intracisternally, intravascularly,intraventricularly, intraosseously, via irrigation of infected bone, orvia application as part of any admixture with a prosthetic devices. Insome embodiments, the administration method includes oral or parenteraladministration.

Compounds provided herein intended for pharmaceutical use may beadministered as crystalline or amorphous products. Pharmaceuticallyacceptable compositions may include solid, semi-solid, liquid,solutions, colloidal, liposomes, emulsions, suspensions, complexes,coacervates and aerosols. Dosage forms, such as, e.g., tablets,capsules, powders, liquids, suspensions, suppositories, aerosols,implants, controlled release or the like. They may be obtained, forexample, as solid plugs, powders, or films by methods such asprecipitation, crystallization, milling, grinding, supercritical fluidprocessing, coacervation, complex coacervation, encapsulation,emulsification, complexation, freeze drying, spray drying, orevaporative drying. Microwave or radio frequency drying may be used forthis purpose. The compounds can also be administered in sustained orcontrolled release dosage forms, including depot injections, osmoticpumps, pills (tablets and or capsules), transdermal (includingelectrotransport) patches, implants and the like, for prolonged and/ortimed, pulsed administration at a predetermined rate.

The compounds can be administered either alone or in combination with aconventional pharmaceutical carrier, excipient or the like.Pharmaceutically acceptable excipients include, but are not limited to,ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifyingdrug delivery systems (SEDDS) such as d-α-tocopherol polyethylene glycol1000 succinate, surfactants used in pharmaceutical dosage forms such asTweens, poloxamers or other similar polymeric delivery matrices, serumproteins, such as human serum albumin, buffer substances such asphosphates, tris, glycine, sorbic acid, potassium sorbate, partialglyceride mixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium-chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethyl cellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, andwool fat. Cyclodextrins such as α-, β, and γ-cyclodextrin, or chemicallymodified derivatives such as hydroxyalkylcyclodextrins, including 2- and3-hydroxypropyl-β-cyclodextrins, or other solubilized derivatives canalso be used to enhance delivery of compounds described herein. Dosageforms or compositions containing a compound as described herein in therange of 0.005% to 100% with the balance made up from non-toxic carriermay be prepared. The contemplated compositions may contain 0.001%-100%of a compound provided herein, in one embodiment 0.1-95%, in anotherembodiment 75-85%, in a further embodiment 20-80%. Actual methods ofpreparing such dosage forms are known, or will be apparent, to thoseskilled in this art; for example, see Remington: The Science andPractice of Pharmacy, 22^(nd) Edition (Pharmaceutical Press, London, UK. 2012).

In one embodiment, the compositions will take the form of a unit dosageform such as a pill or tablet and thus the composition may contain,along with a compound provided herein, a diluent such as lactose,sucrose, dicalcium phosphate, or the like; a lubricant such as magnesiumstearate or the like; and a binder such as starch, gum acacia,polyvinylpyrrolidine, gelatin, cellulose, cellulose derivatives or thelike. In another solid dosage form, a powder, marume, solution orsuspension (e.g., in propylene carbonate, vegetable oils, PEG's,poloxamer 124 or triglycerides) is encapsulated in a capsule (gelatin orcellulose base capsule). Unit dosage forms in which one or morecompounds provided herein or additional active agents are physicallyseparated are also contemplated; e.g., capsules with granules (ortablets in a capsule) of each drug; two-layer tablets; two-compartmentgel caps, etc. Enteric coated or delayed release oral dosage forms arealso contemplated.

Liquid pharmaceutically administrable compositions can, for example, beprepared by dissolving, dispersing, etc. a compound provided herein andoptional pharmaceutical adjuvants in a carrier (e.g., water, saline,aqueous dextrose, glycerol, glycols, ethanol or the like) to form asolution, colloid, liposome, emulsion, complexes, coacervate orsuspension. If desired, the pharmaceutical composition can also containminor amounts of nontoxic auxiliary substances such as wetting agents,emulsifying agents, co-solvents, solubilizing agents, pH bufferingagents and the like (e.g., sodium acetate, sodium citrate, cyclodextrinderivatives, sorbitan monolaurate, triethanolamine acetate,triethanolamine oleate, and the like).

In some embodiments, the unit dosage of compounds of Formula (I) isabout 0.25 mg/Kg to about 50 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 0.25 mg/Kg to about 20 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 0.50 mg/Kg to about 19 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 0.75 mg/Kg to about 18 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 1.0 mg/Kg to about 17 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 1.25 mg/Kg to about 16 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 1.50 mg/Kg to about 15 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 1.75 mg/Kg to about 14 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 2.0 mg/Kg to about 13 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 3.0 mg/Kg to about 12 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 4.0 mg/Kg to about 11 mg/Kg in humans.

In some embodiments, the unit dosage of compounds of Formula (I) isabout 5.0 mg/Kg to about 10 mg/Kg in humans.

In some embodiments, the compositions are provided in unit dosage formssuitable for single administration.

In some embodiments, the compositions are provided in unit dosage formssuitable for twice a day administration.

In some embodiments, the compositions are provided in unit dosage formssuitable for three times a day administration.

Injectables can be prepared in conventional forms, either as liquidsolutions, colloid, liposomes, complexes, coacervate or suspensions, asemulsions, or in solid forms suitable for reconstitution in liquid priorto injection. The percentage of a compound provided herein contained insuch parenteral compositions is highly dependent on the specific naturethereof, as well as the activity of the compound and the needs of thepatient. However, percentages of active ingredient of 0.01% to 10% insolution are employable, and could be higher if the composition is asolid or suspension, which could be subsequently diluted to the abovepercentages.

In some embodiments, the composition will comprise about 0.1-10% of theactive agent in solution.

In some embodiments, the composition will comprise about 0.1-5% of theactive agent in solution.

In some embodiments, the composition will comprise about 0.1-4% of theactive agent in solution.

In some embodiments, the composition will comprise about 0.15-3% of theactive agent in solution.

In some embodiments, the composition will comprise about 0.2-2% of theactive agent in solution.

In some embodiments, the compositions are provided in dosage formssuitable for continuous dosage by intravenous infusion over a period ofabout 1-96 hours.

In some embodiments, the compositions are provided in dosage formssuitable for continuous dosage by intravenous infusion over a period ofabout 1-72 hours.

In some embodiments, the compositions are provided in dosage formssuitable for continuous dosage by intravenous infusion over a period ofabout 1-48 hours.

In some embodiments, the compositions are provided in dosage formssuitable for continuous dosage by intravenous infusion over a period ofabout 1-24 hours.

In some embodiments, the compositions are provided in dosage formssuitable for continuous dosage by intravenous infusion over a period ofabout 1-12 hours.

In some embodiments, the compositions are provided in dosage formssuitable for continuous dosage by intravenous infusion over a period ofabout 1-6 hours.

In some embodiments, these compositions can be administered byintravenous infusion to humans at doses of about 5 mg/m² to about 300mg/m².

In some embodiments, these compositions can be administered byintravenous infusion to humans at doses of about 5 mg/m² to about 200mg/m².

In some embodiments, these compositions can be administered byintravenous infusion to humans at doses of about 5 mg/m² to about 100mg/m².

In some embodiments, these compositions can be administered byintravenous infusion to humans at doses of about 10 mg/m² to about 50mg/m².

In some embodiments, these compositions can be administered byintravenous infusion to humans at doses of about 50 mg/m² to about 200mg/m².

In some embodiments, these compositions can be administered byintravenous infusion to humans at doses of about 75 mg/m² to about 175mg/m².

In some embodiments, these compositions can be administered byintravenous infusion to humans at doses of about 100 mg/m² to about 150mg/m².

It is to be noted that concentrations and dosage values may also varydepending on the specific compound and the severity of the condition tobe alleviated. It is to be further understood that for any particularpatient, specific dosage regimens should be adjusted over time accordingto the individual need and the professional judgment of the personadministering or supervising the administration of the compositions, andthat the concentration ranges set forth herein are exemplary only andare not intended to limit the scope or practice of the claimedcompositions.

In one embodiment, the compositions can be administered to therespiratory tract (including nasal and pulmonary) e.g., through anebulizer, metered-dose inhalers, atomizer, mister, aerosol, dry powderinhaler, insufflator, liquid instillation or other suitable device ortechnique.

In some embodiments, aerosols intended for delivery to the nasal mucosaare provided for inhalation through the nose. For optimal delivery tothe nasal cavities, inhaled particle sizes of about 5 to about 100microns are useful, with particle sizes of about 10 to about 60 micronsbeing preferred. For nasal delivery, a larger inhaled particle size maybe desired to maximize impaction on the nasal mucosa and to minimize orprevent pulmonary deposition of the administered formulation. In someembodiments, aerosols intended for delivery to the lung are provided forinhalation through the nose or the mouth. For delivery to the lung,inhaled aerodynamic particle sizes of about less than 10 μm are useful(e.g., about 1 to about 10 microns). Inhaled particles may be defined asliquid droplets containing dissolved drug, liquid droplets containingsuspended drug particles (in cases where the drug is insoluble in thesuspending medium), dry particles of pure drug substance, drug substanceincorporated with excipients, liposomes, emulsions, colloidal systems,coacervates, aggregates of drug nanoparticles, or dry particles of adiluent which contain embedded drug nanoparticles.

In some embodiments, compounds of Formula (I) disclosed herein intendedfor respiratory delivery (either systemic or local) can be administeredas aqueous formulations, as non-aqueous solutions or suspensions, assuspensions or solutions in halogenated hydrocarbon propellants with orwithout alcohol, as a colloidal system, as emulsions, coacervates, or asdry powders. Aqueous formulations may be aerosolized by liquidnebulizers employing either hydraulic or ultrasonic atomization or bymodified micropump systems (like the soft mist inhalers, the Aerodose®or the AERx® systems). Propellant-based systems may use suitablepressurized metered-dose inhalers (pMDIs). Dry powders may use drypowder inhaler devices (DPIs), which are capable of dispersing the drugsubstance effectively. A desired particle size and distribution may beobtained by choosing an appropriate device.

In some embodiments, the compositions of Formula (I) disclosed hereincan be administered to the ear by various methods. For example, a roundwindow catheter (e.g., U.S. Pat. Nos. 6,440,102 and 6,648,873) can beused.

Alternatively, formulations can be incorporated into a wick for usebetween the outer and middle ear (e.g., U.S. Pat. No. 6,120,484) orabsorbed to collagen sponge or other solid support (e.g., U.S. Pat. No.4,164,559).

If desired, formulations of the disclosure can be incorporated into agel formulation (e.g., U.S. Pat. Nos. 4,474,752 and 6,911,211).

In some embodiments, compounds of Formula (I) disclosed herein intendedfor delivery to the ear can be administered via an implanted pump anddelivery system through a needle directly into the middle or inner ear(cochlea) or through a cochlear implant stylet electrode channel oralternative prepared drug delivery channel such as but not limited to aneedle through temporal bone into the cochlea.

Other options include delivery via a pump through a thin film coatedonto a multichannel electrode or electrode with a specially imbeddeddrug delivery channel (pathways) carved into the thin film for thispurpose. In other embodiments the acidic or basic solid compound ofFormula (I) can be delivered from the reservoir of an external orinternal implanted pumping system.

Formulations of the disclosure also can be administered to the ear byintratympanic injection into the middle ear, inner ear, or cochlea(e.g., U.S. Pat. No. 6,377,849 and Ser. No. 11/337,815).

Intratympanic injection of therapeutic agents is the technique ofinjecting a therapeutic agent behind the tympanic membrane into themiddle and/or inner ear. In one embodiment, the formulations describedherein are administered directly onto the round window membrane viatranstympanic injection. In another embodiment, the ion channelmodulating agent auris-acceptable formulations described herein areadministered onto the round window membrane via a non-transtympanicapproach to the inner ear. In additional embodiments, the formulationdescribed herein is administered onto the round window membrane via asurgical approach to the round window membrane comprising modificationof the crista fenestrae cochleae.

In some embodiments, the compounds of Formula (I) are formulated inrectal compositions such as enemas, rectal gels, rectal foams, rectalaerosols, suppositories, jelly suppositories, or retention enemas,containing conventional suppository bases such as cocoa butter or otherglycerides, as well as synthetic polymers such as polyvinylpyrrolidone,PEG (like PEG ointments), and the like.

Suppositories for rectal administration of the drug (either as asolution, colloid, suspension or a complex) can be prepared by mixing acompound provided herein with a suitable non-irritating excipient thatis solid at ordinary temperatures but liquid at the rectal temperatureand will therefore melt or erode/dissolve in the rectum and release thecompound. Such materials include cocoa butter, glycerinated gelatin,hydrogenated vegetable oils, poloxamers, mixtures of polyethyleneglycols of various molecular weights and fatty acid esters ofpolyethylene glycol. In suppository forms of the compositions, alow-melting wax such as, but not limited to, a mixture of fatty acidglycerides, optionally in combination with cocoa butter, is firstmelted.

Solid compositions can be provided in various different types of dosageforms, depending on the physicochemical properties of the compoundprovided herein, the desired dissolution rate, cost considerations, andother criteria. In one of the embodiments, the solid composition is asingle unit. This implies that one unit dose of the compound iscomprised in a single, physically shaped solid form or article. In otherwords, the solid composition is coherent, which is in contrast to amultiple unit dosage form, in which the units are incoherent.

Examples of single units which may be used as dosage forms for the solidcomposition include tablets, such as compressed tablets, film-likeunits, foil-like units, wafers, lyophilized matrix units, and the like.In one embodiment, the solid composition is a highly porous lyophilizedform. Such lyophilizates, sometimes also called wafers or lyophilizedtablets, are particularly useful for their rapid disintegration, whichalso enables the rapid dissolution of the compound.

On the other hand, for some applications the solid composition may alsobe formed as a multiple unit dosage form as defined above. Examples ofmultiple units are powders, granules, microparticles, pellets,mini-tablets, beads, lyophilized powders, and the like. In oneembodiment, the solid composition is a lyophilized powder. Such adispersed lyophilized system comprises a multitude of powder particles,and due to the lyophilization process used in the formation of thepowder, each particle has an irregular, porous microstructure throughwhich the powder is capable of absorbing water very rapidly, resultingin quick dissolution. Effervescent compositions are also contemplated toaid the quick dispersion and absorption of the compound.

Another type of multiparticulate system which is also capable ofachieving rapid drug dissolution is that of powders, granules, orpellets from water-soluble excipients which are coated with a compoundprovided herein so that the compound is located at the outer surface ofthe individual particles. In this type of system, the water-soluble lowmolecular weight excipient may be useful for preparing the cores of suchcoated particles, which can be subsequently coated with a coatingcomposition comprising the compound and, for example, one or moreadditional excipients, such as a binder, a pore former, a saccharide, asugar alcohol, a film-forming polymer, a plasticizer, or otherexcipients used in pharmaceutical coating compositions.

Also provided herein are kits. Typically, a kit includes one or morecompounds or compositions as described herein. In certain embodiments, akit can include one or more delivery systems, e.g., for delivering oradministering a compound as provided herein, and directions for use ofthe kit (e.g., instructions for treating a patient). In anotherembodiment, the kit can include a compound or composition as describedherein and a label that indicates that the contents are to beadministered to a patient with cancer. In another embodiment, the kitcan include a compound or composition as described herein and a labelthat indicates that the contents are to be administered to a patientwith one or more of hepatocellular carcinoma, colon cancer, leukemia,lymphoma, sarcoma, ovarian cancer, diabetic retinopathy, pulmonaryfibrosis, rheumatoid arthritis, sepsis, ankylosing spondylitis,psoriasis, scleroderma, mycotic and viral infections, bone and cartilagediseases, Alzheimer's disease, lung disease, bone/osteoporotic (wrist,spine, shoulder and hip) fractures, articular cartilage (chondral)defects, degenerative disc disease (or intervertebral discdegeneration), polyposis coli, bone density and vascular defects in theeye (Osteoporosis-pseudoglioma Syndrome, OPPG), familial exudativevitreoretinopathy, retinal angiogenesis, early coronary disease,tetra-amelia, Müllerian-duct regression and virilization, SERKALsyndrome, type II diabetes, Fuhrmann syndrome,Al-Awadi/Raas-Rothschild/Schinzel phocomelia syndrome,odonto-onycho-dermal dysplasia, obesity, split-hand/foot malformation,caudal duplication, tooth agenesis, Wilms tumor, skeletal dysplasia,focal dermal hypoplasia, autosomal recessive anonychia, neural tubedefects, alpha-thalassemia (ATRX) syndrome, fragile X syndrome, ICFsyndrome, Angelman syndrome, Prader-Willi syndrome, Beckwith-WiedemannSyndrome, Norrie disease, and Rett syndrome.

Methods of Treatment

The compounds and compositions provided herein can be used as inhibitorsand/or modulators of one or more components of the Wnt pathway, whichmay include one or more Wnt proteins, and thus can be used to treat avariety of disorders and diseases in which aberrant Wnt signaling isimplicated, such as cancer and other diseases associated with abnormalangiogenesis, cellular proliferation, and cell cycling. Accordingly, thecompounds and compositions provided herein can be used to treat cancer,to reduce or inhibit angiogenesis, to reduce or inhibit cellularproliferation, to correct a genetic disorder, and/or to treat aneurological condition/disorder/disease due to mutations ordysregulation of the Wnt pathway and/or of one or more of Wnt signalingcomponents. Non-limiting examples of diseases which can be treated withthe compounds and compositions provided herein include a variety ofcancers, diabetic retinopathy, pulmonary fibrosis, rheumatoid arthritis,scleroderma, mycotic and viral infections, bone and cartilage diseases,neurological conditions/diseases such as Alzheimer's disease,amyotrophic lateral sclerosis (ALS), motor neuron disease, multiplesclerosis or autism, lung disease, bone/osteoporotic (wrist, spine,shoulder and hip) fractures, polyposis coli, bone density and vasculardefects in the eye (Osteoporosis-pseudoglioma Syndrome, OPPG), familialexudative vitreoretinopathy, retinal angiogenesis, early coronarydisease, tetra-amelia, Müllerian-duct regression and virilization,SERKAL syndrome, type II diabetes, Fuhrmann syndrome,Al-Awadi/Raas-Rothschild/Schinzel phocomelia syndrome,odonto-onycho-dermal dysplasia, obesity, split-hand/foot malformation,caudal duplication, tooth agenesis, Wilms tumor, skeletal dysplasia,focal dermal hypoplasia, autosomal recessive anonychia, neural tubedefects, alpha-thalassemia (ATRX) syndrome, fragile X syndrome, ICFsyndrome, Angelman syndrome, Prader-Willi syndrome, Beckwith-WiedemannSyndrome, Norrie disease and Rett syndrome.

With respect to cancer, the Wnt pathway is known to be constitutivelyactivated in a variety of cancers including, for example, colon cancer,hepatocellular carcinoma, lung cancer, ovarian cancer, prostate cancer,pancreatic cancer and leukemias such as CML, CLL and T-ALL. Accordingly,the compounds and compositions described herein may be used to treatthese cancers in which the Wnt pathway is constitutively activated. Incertain embodiments, the cancer is chosen from hepatocellular carcinoma,colon cancer, leukemia, lymphoma, sarcoma and ovarian cancer.

Other cancers can also be treated with the compounds and compositionsdescribed herein.

More particularly, cancers that may be treated by the compounds,compositions and methods described herein include, but are not limitedto, the following:

1) Breast cancers, including, for example ER⁺ breast cancer, ER⁻ breastcancer, her2⁻ breast cancer, her2⁺ breast cancer, stromal tumors such asfibroadenomas, phyllodes tumors, and sarcomas, and epithelial tumorssuch as large duct papillomas; carcinomas of the breast including insitu (noninvasive) carcinoma that includes ductal carcinoma in situ(including Paget's disease) and lobular carcinoma in situ, and invasive(infiltrating) carcinoma including, but not limited to, invasive ductalcarcinoma, invasive lobular carcinoma, medullary carcinoma, colloid(mucinous) carcinoma, tubular carcinoma, and invasive papillarycarcinoma; and miscellaneous malignant neoplasms. Further examples ofbreast cancers can include luminal A, luminal B, basal A, basal B, andtriple negative breast cancer, which is estrogen receptor negative(ER⁻), progesterone receptor negative, and her2 negative (her2⁻). Insome embodiments, the breast cancer may have a high risk Oncotype score.

2) Cardiac cancers, including, for example sarcoma, e.g., angiosarcoma,fibrosarcoma, rhabdomyosarcoma, and liposarcoma; myxoma; rhabdomyoma;fibroma; lipoma and teratoma.

3) Lung cancers, including, for example, bronchogenic carcinoma, e.g.,squamous cell, undifferentiated small cell, undifferentiated large cell,and adenocarcinoma; alveolar and bronchiolar carcinoma; bronchialadenoma; sarcoma; lymphoma; chondromatous hamartoma; and mesothelioma.

4) Gastrointestinal cancer, including, for example, cancers of theesophagus, e.g., squamous cell carcinoma, adenocarcinoma,leiomyosarcoma, and lymphoma; cancers of the stomach, e.g., carcinoma,lymphoma, and leiomyosarcoma; cancers of the pancreas, e.g., ductaladenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors,and vipoma; cancers of the small bowel, e.g., adenocarcinoma, lymphoma,carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma,neurofibroma, and fibroma; cancers of the large bowel, e.g.,adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, andleiomyoma.

5) Genitourinary tract cancers, including, for example, cancers of thekidney, e.g., adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma,and leukemia; cancers of the bladder and urethra, e.g., squamous cellcarcinoma, transitional cell carcinoma, and adenocarcinoma; cancers ofthe prostate, e.g., adenocarcinoma, and sarcoma; cancer of the testis,e.g., seminoma, teratoma, embryonal carcinoma, teratocarcinoma,choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma,fibroadenoma, adenomatoid tumors, and lipoma.

6) Liver cancers, including, for example, hepatoma, e.g., hepatocellularcarcinoma; cholangiocarcinoma; hepatoblastoma; angiosarcoma;hepatocellular adenoma; and hemangioma.

7) Bone cancers, including, for example, osteogenic sarcoma(osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma,chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cellsarcoma), multiple myeloma, malignant giant cell tumor chordoma,osteochrondroma (osteocartilaginous exostoses), benign chondroma,chondroblastoma, chondromyxofibroma, osteoid osteoma and giant celltumors.

8) Nervous system cancers, including, for example, cancers of the skull,e.g., osteoma, hemangioma, granuloma, xanthoma, and osteitis deformans;cancers of the meninges, e.g., meningioma, meningiosarcoma, andgliomatosis; cancers of the brain, e.g., astrocytoma, medulloblastoma,glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform,oligodendroglioma, oligodendrocytoma, schwannoma, retinoblastoma, andcongenital tumors; and cancers of the spinal cord, e.g., neurofibroma,meningioma, glioma, and sarcoma.

9) Gynecological cancers, including, for example, cancers of the uterus,e.g., endometrial carcinoma; cancers of the cervix, e.g., cervicalcarcinoma, and pre tumor cervical dysplasia; cancers of the ovaries,e.g., ovarian carcinoma, including serous cystadenocarcinoma, mucinouscystadenocarcinoma, unclassified carcinoma, granulosa theca cell tumors,Sertoli Leydig cell tumors, dysgerminoma, and malignant teratoma;cancers of the vulva, e.g., squamous cell carcinoma, intraepithelialcarcinoma, adenocarcinoma, fibrosarcoma, and melanoma; cancers of thevagina, e.g., clear cell carcinoma, squamous cell carcinoma, botryoidsarcoma, and embryonal rhabdomyosarcoma; and cancers of the fallopiantubes, e.g., carcinoma.

10) Hematologic cancers, including, for example, cancers of the blood,e.g., acute myeloid leukemia, chronic myeloid leukemia, acutelymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferativediseases, multiple myeloma, and myelodysplastic syndrome, Hodgkin'slymphoma, non-Hodgkin's lymphoma (malignant lymphoma) and Waldenström'smacroglobulinemia.

11) Skin cancers and skin disorders, including, for example, malignantmelanoma and metastatic melanoma, basal cell carcinoma, squamous cellcarcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, angioma,dermatofibroma, keloids, and scleroderma.

12) Adrenal gland cancers, including, for example, neuroblastoma.

More particularly, tumors of the central nervous system that may betreated by the compounds, compositions and methods described hereininclude:

1) Astrocytic tumors, e.g., diffuse astrocytoma (fibrillary,protoplasmic, gemistocytic, mixed), anaplastic (malignant) astrocytoma,glioblastoma multiforme (giant cell glioblastoma and gliosarcoma),pilocytic astrocytoma (pilomyxoid astrocytoma), pleomorphicxanthoastrocytoma, subependymal giant cell astrocytoma, and gliomatosiscerebri.

2) Oligodendroglial tumors, e.g., oligodendroglioma and anaplasticoligodendroglioma.

3) Oligoastrocytic tumors, e.g., oligoastrocytoma and anaplasticoligoastrocytoma.

4) Ependymal tumors, e.g., subependymoma, myxopapillary ependymoma,ependymoma, (cellular, papillary, clear cell, tanycytic), and anaplastic(malignant) ependymoma.

5) Choroid plexus tumors, e.g., choroid plexus papilloma, atypicalchoroid plexus papilloma, and choroid plexus carcinoma.

6) Neuronal and mixed neuronal-glial tumors, e.g., gangliocytoma,ganglioglioma, dysembryoplastic neuroepithelial tumor (DNET), dysplasticgangliocytoma of the cerebellum (Lhermitte-Duclos), desmoplasticinfantile astrocytoma/ganglioglioma, central neurocytoma, anaplasticganglioglioma, extraventricular neurocytoma, cerebellar liponeurocytoma,Papillary glioneuronal tumor, Rosette-forming glioneuronal tumor of thefourth ventricle, and paraganglioma of the filum terminale.

7) Pineal tumors, e.g., pineocytoma, pineoblastoma, papillary tumors ofthe pineal region, and pineal parenchymal tumor of intermediatedifferentiation.

8) Embryonal tumors, e.g., medulloblastoma (medulloblastoma withextensive nodularity, anaplastic medulloblastoma, desmoplastic, largecell, melanotic, medullomyoblastoma), medulloepithelioma, supratentorialprimitive neuroectodermal tumors, and primitive neuroectodermal tumors(PNETs) such as neuroblastoma, ganglioneuroblastoma, ependymoblastoma,and atypical teratoid/rhabdoid tumor.

9) Neuroblastic tumors, e.g., olfactory (esthesioneuroblastoma),olfactory neuroepithelioma, and neuroblastomas of the adrenal gland andsympathetic nervous system.

10) Glial tumors, e.g., astroblastoma, chordoid glioma of the thirdventricle, and angiocentric glioma.

11) Tumors of cranial and paraspinal nerves, e.g., schwannoma,neurofibroma Perineurioma, and malignant peripheral nerve sheath tumor.

12) Tumors of the meninges such as tumors of meningothelial cells, e.g.,meningioma (atypical meningioma and anaplastic meningioma); mesenchymaltumors, e.g., lipoma, angiolipoma, hibernoma, liposarcoma, solitaryfibrous tumor, fibrosarcoma, malignant fibrous histiocytoma, leiomyoma,leiomyosarcoma, rhabdomyoma, rhabdomyosarcoma, chondroma,chondrosarcoma, osteoma, osteosarcoma, osteochondroma, haemangioma,epithelioid hemangioendothelioma, haemangiopericytoma, anaplastichaemangiopericytoma, angiosarcoma, Kaposi Sarcoma, and Ewing Sarcoma;primary melanocytic lesions, e.g., diffuse melanocytosis, melanocytoma,malignant melanoma, meningeal melanomatosis; and hemangioblastomas.

13) Tumors of the hematopoietic system, e.g., malignant Lymphomas,plasmocytoma, and granulocytic sarcoma.

14) Germ cell tumors, e.g., germinoma, embryonal carcinoma, yolk sactumor, choriocarcinoma, teratoma, and mixed germ cell tumors.

15) Tumors of the sellar region, e.g., craniopharyngioma, granular celltumor, pituicytoma, and spindle cell oncocytoma of the adenohypophysis.

Cancers may be solid tumors that may or may not be metastatic. Cancersmay also occur, as in leukemia, as a diffuse tissue. Thus, the term“tumor cell,” as provided herein, includes a cell afflicted by any oneof the above identified disorders.

A method of treating cancer using a compound or composition as describedherein may be combined with existing methods of treating cancers, forexample by chemotherapy, irradiation, or surgery (e.g., oophorectomy).In some embodiments, a compound or composition can be administeredbefore, during, or after another anticancer agent or treatment.

The compounds and compositions described herein can be used asanti-angiogenesis agents and as agents for modulating and/or inhibitingthe activity of protein kinases, thus providing treatments for cancerand other diseases associated with cellular proliferation mediated byprotein kinases. For example, the compounds described herein can inhibitthe activity of one or more kinases. Accordingly, provided herein is amethod of treating cancer or preventing or reducing angiogenesis throughkinase inhibition.

In addition, and including treatment of cancer, the compounds andcompositions described herein can function as cell-cycle control agentsfor treating proliferative disorders in a patient. Disorders associatedwith excessive proliferation include, for example, cancers, scleroderma,immunological disorders involving undesired proliferation of leukocytes,and restenosis and other smooth muscle disorders. Furthermore, suchcompounds may be used to prevent de-differentiation of post-mitotictissue and/or cells.

Diseases or disorders associated with uncontrolled or abnormal cellularproliferation include, but are not limited to, the following:

-   -   a variety of cancers, including, but not limited to, carcinoma,        hematopoietic tumors of lymphoid lineage, hematopoietic tumors        of myeloid lineage, tumors of mesenchymal origin, tumors of the        central and peripheral nervous system and other tumors including        melanoma, seminoma and Kaposi's sarcoma.    -   a disease process which features abnormal cellular        proliferation, e.g., benign prostatic hyperplasia, familial        adenomatosis polyposis, neurofibromatosis, atherosclerosis,        arthritis, glomerulonephritis, restenosis following angioplasty        or vascular surgery, inflammatory bowel disease, transplantation        rejection, endotoxic shock, and fungal infections. Fibrotic        disorders such as skin fibrosis; scleroderma; progressive        systemic fibrosis; lung fibrosis; muscle fibrosis; kidney        fibrosis; glomerulosclerosis; glomerulonephritis; hypertrophic        scar formation; uterine fibrosis; renal fibrosis; cirrhosis of        the liver, liver fibrosis; fatty liver disease (FLD); adhesions,        such as those occurring in the abdomen, pelvis, spine or        tendons; chronic obstructive pulmonary disease; fibrosis        following myocardial infarction; pulmonary fibrosis; fibrosis        and scarring associated with diffuse/interstitial lung disease;        central nervous system fibrosis, such as fibrosis following        stroke; fibrosis associated with neuro-degenerative disorders        such as Alzheimer's Disease or multiple sclerosis; fibrosis        associated with proliferative vitreoretinopathy (PVR);        restenosis; endometriosis; ischemic disease and radiation        fibrosis.    -   defective apoptosis-associated conditions, such as cancers        (including but not limited to those types mentioned herein),        viral infections (including but not limited to herpesvirus,        poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus),        prevention of AIDS development in HIV-infected individuals,        autoimmune diseases (including but not limited to systemic lupus        erythematosus, rheumatoid arthritis, sepsis, ankylosing        spondylitis, psoriasis, scleroderma, autoimmune mediated        glomerulonephritis, inflammatory bowel disease and autoimmune        diabetes mellitus), neuro-degenerative disorders (including but        not limited to Alzheimer's disease, lung disease, amyotrophic        lateral sclerosis, retinitis pigmentosa, Parkinson's disease,        AIDS-related dementia, spinal muscular atrophy and cerebellar        degeneration), myelodysplastic syndromes, aplastic anemia,        ischemic injury associated with myocardial infarctions, stroke        and reperfusion injury, arrhythmia, atherosclerosis,        toxin-induced or alcohol related liver diseases, hematological        diseases (including but not limited to chronic anemia and        aplastic anemia), degenerative diseases of the musculoskeletal        system (including but not limited to osteoporosis and        arthritis), tendinopathies such as tendinitis and tendinosis,        aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple        sclerosis, kidney diseases and cancer pain.    -   genetic diseases due to mutations in Wnt signaling components,        such as polyposis coli, bone density and vascular defects in the        eye (Osteoporosis-pseudoglioma Syndrome, OPPG), familial        exudative vitreoretinopathy, retinal angiogenesis, early        coronary disease, tetra-amelia, Müllerian-duct regression and        virilization, SERKAL syndrome, type II diabetes, Fuhrmann        syndrome, Al-Awadi/Raas-Rothschild/Schinzel phocomelia syndrome,        odonto-onycho-dermal dysplasia, obesity, split-hand/foot        malformation, caudal duplication, tooth agenesis, Wilms tumor,        skeletal dysplasia, focal dermal hypoplasia, autosomal recessive        anonychia, neural tube defects, alpha-thalassemia (ATRX)        syndrome, fragile X syndrome, ICF syndrome, Angelman syndrome,        Prader-Willi syndrome, Beckwith-Wiedemann Syndrome, Norrie        disease and Rett syndrome.

The compounds and compositions provided herein have been found topossess immunomodulatory activities and are expected to control theinnate and adaptive immune system (e.g. macrophages, microglia,dendritic cells, B and T cells) and suppress pro-inflammatory cytokinerelease (e.g. TNF, IL-6, IL-1, IFN□) which is well known to be involvedin chronic inflammation in a wide variety of disease areas. Thereforecompounds and compositions provided herein can used to treat chronicinflammation associated with disorders and diseases including but notlimited to eye disorders, joint pain, arthritis (rheumatoid, osteo,psoriatic gout), cancers (colon, breast, lung, pancreas, and others),gastrointestinal disorders (ulcerative colitis and inflammatory boweldiseases), pulmonary disorders (chronic obstructive pulmonary disorderand asthma), allergies, skin disorders (atopic dermatitis andpsoriasis), diabetes, pancreatitis, tendonitis, hepatitis, heartdisease, myocarditis, stroke, lupus, and neurological disorders such asmultiple sclerosis, Parkinson's and dementia including Alzheimer'sdisease.

The compounds and compositions provided herein can be used as inhibitorsand/or modulators of the enzyme DYRK1A, and thus can be used to treat avariety of disorders and diseases associated with tau protein, amyloid,alpha-synuclein, TDP-43 or FUS pathology including, but not limited to,Alzheimer's disease, amyotrophic lateral sclerosis (ALS), down syndrome,frontotemporal dementia (FTD) including FTD with Parkinsonism-17(FTDP-17), behavioural variant frontotemporal dementia (bvFTD), FTD inpatients with motor neuron disease (MND) (typically amyotrophic lateralsclerosis, also called FTD-ALS), corticobasal degeneration (CBD) (alsocalled corticobasal ganglionic degeneration), progressive supranuclearpalsy, primary progressive aphasia (PPA), globular glial tauopathy(GGT), myotonic dystrophy type 1 (DM1) (also called Steinert disease),myotonic dystrophy type 2 (DM2) (also called proximal myotonicmyopathy), Guam complex, argyrophilic grain disease, dementiapugilistica, post-encephalitic parkinsonism, Lewy body dementia,Parkinson's disease, Pick's disease, and additional diseases withpronounced neurodegeneration such as autism, dementia, epilepsy,Huntington's disease, multiple sclerosis; diseases and disordersassociated with acquired brain injury such as chronic traumaticencephalopathy, traumatic brain injury, tumor, and stroke.

Non-limiting examples of neurological disorders (e.g., neurologicalconditions and neurological diseases) which can be treated with thecompounds and compositions provided herein include Alzheimer's disease,aphasia, apraxia, arachnoiditis, ataxia telangiectasia, attentiondeficit hyperactivity disorder, auditory processing disorder, autism,alcoholism, Bell's palsy, bipolar disorder, brachial plexus injury,Canavan disease, carpal tunnel syndrome, causalgia, central painsyndrome, central pontine myelinolysis, centronuclear myopathy, cephalicdisorder, cerebral aneurysm, cerebral arteriosclerosis, cerebralatrophy, cerebral gigantism, cerebral palsy, cerebral vasculitis,cervical spinal stenosis, Charcot-Marie-Tooth disease, Chiarimalformation, chronic fatigue syndrome, chronic inflammatorydemyelinating polyneuropathy (CIDP), chronic pain, Coffin-Lowrysyndrome, complex regional pain syndrome, compression neuropathy,congenital facial diplegia, corticobasal degeneration, cranialarteritis, craniosynostosis, Creutzfeldt-Jakob disease, cumulativetrauma disorder, Cushing's syndrome, cytomegalic inclusion body disease(CIBD), Dandy-Walker syndrome, Dawson disease, De Morsier's syndrome,Dejerine-Klumpke palsy, Dejerine-Sottas disease, delayed sleep phasesyndrome, dementia, dermatomyositis, developmental dyspraxia, diabeticneuropathy, diffuse sclerosis, Dravet syndrome, dysautonomia,dyscalculia, dysgraphia, dyslexia, dystonia, empty sella syndrome,encephalitis, encephalocele, encephalotrigeminal angiomatosis,encopresis, epilepsy, Erb's palsy, erythromelalgia, essential tremor,Fabry's disease, Fahr's syndrome, familial spastic paralysis, febrileseizure, Fisher syndrome, Friedreich's ataxia, fibromyalgia, Foville'ssyndrome, Gaucher's disease, Gerstmann's syndrome, giant cell arteritis,giant cell inclusion disease, globoid cell leukodystrophy, gray matterheterotopia, Guillain-Barré syndrome, HTLV-1 associated myelopathy,Hallervorden-Spatz disease, hemifacial spasm, hereditary spasticparaplegia, heredopathia atactica polyneuritiformis, herpes zosteroticus, herpes zoster, Hirayama syndrome, holoprosencephaly,Huntington's disease, hydranencephaly, hydrocephalus, hypercortisolism,hypoxia, immune-mediated encephalomyelitis, inclusion body myositis,incontinentia pigmenti, infantile phytanic acid storage disease,infantile Refsum disease, infantile spasms, inflammatory myopathy,intracranial cyst, intracranial hypertension, Joubert syndrome, Karaksyndrome, Kearns-Sayre syndrome, Kennedy disease, Kinsbourne syndrome,Klippel Feil syndrome, Krabbe disease, Kugelberg-Welander disease, kuru,Lafora disease, Lambert-Eaton myasthenic syndrome, Landau-Kleffnersyndrome, lateral medullary (Wallenberg) syndrome, Leigh's disease,Lennox-Gastaut syndrome, Lesch-Nyhan syndrome, leukodystrophy, Lewy bodydementia, lissencephaly, locked-in syndrome, Lou Gehrig's disease,lumbar disc disease, lumbar spinal stenosis, Lyme disease,Machado-Joseph disease (Spinocerebellar ataxia type 3), macrencephaly,macropsia, megalencephaly, Melkersson-Rosenthal syndrome, Meniere'sdisease, meningitis, Menkes disease, metachromatic leukodystrophy,microcephaly, micropsia, Miller Fisher syndrome, misophonia,mitochondrial myopathy, Mobius syndrome, monomelic amyotrophy, motorneuron disease, motor skills disorder, Moyamoya disease,mucopolysaccharidoses, multi-infarct dementia, multifocal motorneuropathy, multiple sclerosis, multiple system atrophy, musculardystrophy, myalgic encephalomyelitis, myasthenia gravis, myelinoclasticdiffuse sclerosis, myoclonic Encephalopathy of infants, myoclonus,myopathy, myotubular myopathy, myotonia congenital, narcolepsy,neurofibromatosis, neuroleptic malignant syndrome, lupus erythematosus,neuromyotonia, neuronal ceroid lipofuscinosis, Niemann-Pick disease,O'Sullivan-McLeod syndrome, occipital Neuralgia, occult SpinalDysraphism Sequence, Ohtahara syndrome, olivopontocerebellar atrophy,opsoclonus myoclonus syndrome, optic neuritis, orthostatic hypotension,palinopsia, paresthesia, Parkinson's disease, paramyotonia Congenita,paraneoplastic diseases, paroxysmal attacks, Parry-Romberg syndrome,Pelizaeus-Merzbacher disease, periodic paralyses, peripheral neuropathy,photic sneeze reflex, phytanic acid storage disease, Pick's disease,polymicrogyria (PMG), polymyositis, porencephaly, post-polio syndrome,postherpetic neuralgia (PHN), postural hypotension, Prader-Willisyndrome, primary lateral sclerosis, prion diseases, progressivehemifacial atrophy, progressive multifocal leukoencephalopathy,progressive supranuclear palsy, pseudotumor cerebri, Ramsay Huntsyndrome type I, Ramsay Hunt syndrome type II, Ramsay Hunt syndrome typeIII, Rasmussen's encephalitis, reflex neurovascular dystrophy, Refsumdisease, restless legs syndrome, retrovirus-associated myelopathy, Rettsyndrome, Reye's syndrome, rhythmic movement disorder, Romberg syndrome,Saint Vitus dance, Sandhoff disease, schizophrenia, Schilder's disease,schizencephaly, sensory integration dysfunction, septo-optic dysplasia,Shy-Drager syndrome, Sjögren's syndrome, snatiation, Sotos syndrome,spasticity, spina bifida, spinal cord tumors, spinal muscular atrophy,spinocerebellar ataxia, Steele-Richardson-Olszewski syndrome,Stiff-person syndrome, stroke, Sturge-Weber syndrome, subacutesclerosing panencephalitis, subcortical arteriosclerotic encephalopathy,superficial siderosis, Sydenham's chorea, syncope, synesthesia,syringomyelia, tarsal tunnel syndrome, tardive dyskinesia, tardivedysphrenia, Tarlov cyst, Tay-Sachs disease, temporal arteritis, tetanus,tethered spinal cord syndrome, Thomsen disease, thoracic outletsyndrome, tic douloureux, Todd's paralysis, Tourette syndrome, toxicencephalopathy, transient ischemic attack, transmissible spongiformencephalopathies, transverse myelitis, tremor, trigeminal neuralgia,tropical spastic paraparesis, trypanosomiasis, tuberous sclerosis,ubisiosis, Von Hippel-Lindau disease (VHL), Viliuisk Encephalomyelitis(VE), Wallenberg's syndrome, Werdnig, Hoffman disease, west syndrome,Williams syndrome, Wilson's disease, and Zellweger syndrome.

The compounds and compositions may also be useful in the inhibition ofthe development of invasive cancer, tumor angiogenesis and metastasis.

In some embodiments, the disclosure provides a method for treating adisease or disorder associated with aberrant cellular proliferation byadministering to a patient in need of such treatment an effective amountof one or more of the compounds of Formula (I), in combination(simultaneously or sequentially) with at least one other agent.

In some embodiments, the disclosure provides a method of treating orameliorating in a patient a disorder or disease selected from the groupconsisting of: cancer, pulmonary fibrosis, idiopathic pulmonary fibrosis(IPF), degenerative disc disease, bone/osteoporotic fractures, bone orcartilage disease, and osteoarthritis, the method comprisingadministering to the patient a therapeutically effective amount of acompound according to Formula (I), or a pharmaceutically acceptable saltthereof.

In some embodiments, the disclosure provides a method of treating orameliorating in a patient a disorder or disease selected from the groupconsisting of: chronic inflammation, systemic inflammation, diabetes,cancer, pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF),degenerative disc disease, bone/osteoporotic fractures, a bone orcartilage disease, a neurological condition/disorder/disease,osteoarthritis, lung disease, a fibrotic disorder.

In some embodiments, the pharmaceutical composition comprises atherapeutically effective amount of a compound of Formula (I), or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient.

In some embodiments, the method of treats a disorder or disease in whichaberrant Wnt signaling is implicated in a patient, the method comprisesadministering to the patient a therapeutically effective amount of acompound of Formula (I), or a pharmaceutically acceptable salt thereof.

In some embodiments, the disorder or disease is the pain andinflammation associated with cancer.

In some embodiments, the disorder or disease is the pain andinflammation associated with a joint.

In some embodiments, the disorder or disease is the pain andinflammation associated with the knee.

In some embodiments, the disorder or disease is the pain andinflammation associated with the hip.

In some embodiments, the disorder or disease is the pain andinflammation associated with the shoulder.

In some embodiments, the disorder or disease is the pain andinflammation associated with arthritis.

In some embodiments, the disorder or disease is the pain andinflammation associated with gastrointestinal disorders.

In some embodiments, the disorder or disease is the pain andinflammation associated with pulmonary disorders.

In some embodiments, the disorder or disease is the pain andinflammation associated with allergies.

In some embodiments, the disorder or disease is the pain andinflammation associated with skin disorders.

In some embodiments, the disorder or disease is the pain andinflammation associated with diabetes.

In some embodiments, the disorder or disease is the pain andinflammation associated with pancreatitis.

In some embodiments, the disorder or disease is the pain andinflammation associated with tendonitis.

In some embodiments, the disorder or disease is the pain andinflammation associated with heart disease.

In some embodiments, the disorder or disease is the pain andinflammation associated with lupus.

In some embodiments, the disorder or disease is the pain andinflammation associated with a neurological disorder.

In some embodiments, the disorder or disease is the pain andinflammation associated with multiple sclerosis.

In some embodiments, the disorder or disease is the pain andinflammation associated with Parkinson's.

In some embodiments, the disorder or disease is cancer.

In some embodiments, the disorder or disease is systemic inflammation.

In some embodiments, the disorder or disease is metastatic melanoma.

In some embodiments, the disorder or disease is fatty liver disease.

In some embodiments, the disorder or disease is liver fibrosis.

In some embodiments, the disorder or disease is tendon regeneration.

In some embodiments, the disorder or disease is diabetes.

In some embodiments, the disorder or disease is degenerative discdisease.

In some embodiments, the disorder or disease is osteoarthritis.

In some embodiments, the disorder or disease is diabetic retinopathy.

In some embodiments, the disorder or disease is pulmonary fibrosis.

In some embodiments, the disorder or disease is idiopathic pulmonaryfibrosis (IPF).

In some embodiments, the disorder or disease is degenerative discdisease.

In some embodiments, the disorder or disease is rheumatoid arthritis.

In some embodiments, the disorder or disease is scleroderma.

In some embodiments, the disorder or disease is a mycotic or viralinfection.

In some embodiments, the disorder or disease is a bone or cartilagedisease.

In some embodiments, the disorder or disease is a neurological disorder.

In some embodiments, the disorder or disease is Alzheimer's disease.

In some embodiments, the disorder or disease is osteoarthritis.

In some embodiments, the disorder or disease is lung disease.

In some embodiments, the disorder or disease is a genetic disease causedby mutations in Wnt signaling components, wherein the genetic disease isselected from: polyposis coli, osteoporosis-pseudoglioma syndrome,familial exudative vitreoretinopathy, retinal angiogenesis, earlycoronary disease, tetra-amelia syndrome, Müllerian-duct regression andvirilization, SERKAL syndrome, diabetes mellitus type 2, Fuhrmannsyndrome, Al-Awadi/Raas-Rothschild/Schinzel phocomelia syndrome,odonto-onycho-dermal dysplasia, obesity, split-hand/foot malformation,caudal duplication syndrome, tooth agenesis, Wilms tumor, skeletaldysplasia, focal dermal hypoplasia, autosomal recessive anonychia,neural tube defects, alpha-thalassemia (ATRX) syndrome, fragile Xsyndrome, ICF syndrome, Angelman syndrome, Prader-Willi syndrome,Beckwith-Wiedemann Syndrome, Norrie disease and Rett syndrome.

In some embodiments, the patient is a human.

In some embodiments, the cancer is chosen from: hepatocellularcarcinoma, colon cancer, breast cancer, pancreatic cancer, chronicmyeloid leukemia (CML), chronic myelomonocytic leukemia, chroniclymphocytic leukemia (CLL), acute myeloid leukemia, acute lymphocyticleukemia, Hodgkin lymphoma, lymphoma, sarcoma and ovarian cancer.

In some embodiments, the cancer is chosen from: lung cancer—non-smallcell, lung cancer—small cell, multiple myeloma, nasopharyngeal cancer,neuroblastoma, osteosarcoma, penile cancer, pituitary tumors, prostatecancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skincancer—basal and squamous cell, skin cancer—melanoma, small intestinecancer, stomach (gastric) cancers, testicular cancer, thymus cancer,thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer,laryngeal or hypopharyngeal cancer, kidney cancer, Kaposi sarcoma,gestational trophoblastic disease, gastrointestinal stromal tumor,gastrointestinal carcinoid tumor, gallbladder cancer, eye cancer(melanoma and lymphoma), Ewing tumor, esophagus cancer, endometrialcancer, colorectal cancer, cervical cancer, brain or spinal cord tumor,bone metastasis, bone cancer, bladder cancer, bile duct cancer, analcancer and adrenal cortical cancer.

In some embodiments, the cancer is hepatocellular carcinoma.

In some embodiments, the cancer is colon cancer.

In some embodiments, the cancer is colorectal cancer.

In some embodiments, the cancer is breast cancer.

In some embodiments, the cancer is pancreatic cancer.

In some embodiments, the cancer is chronic myeloid leukemia (CML).

In some embodiments, the cancer is chronic myelomonocytic leukemia.

In some embodiments, the cancer is chronic lymphocytic leukemia (CLL).

In some embodiments, the cancer is acute myeloid leukemia.

In some embodiments, the cancer is acute lymphocytic leukemia.

In some embodiments, the cancer is Hodgkin lymphoma.

In some embodiments, the cancer is lymphoma.

In some embodiments, the cancer is sarcoma.

In some embodiments, the cancer is ovarian cancer.

In some embodiments, the cancer is lung cancer—non-small cell.

In some embodiments, the cancer is lung cancer—small cell.

In some embodiments, the cancer is multiple myeloma.

In some embodiments, the cancer is nasopharyngeal cancer.

In some embodiments, the cancer is neuroblastoma.

In some embodiments, the cancer is osteosarcoma.

In some embodiments, the cancer is penile cancer.

In some embodiments, the cancer is pituitary tumors.

In some embodiments, the cancer is prostate cancer.

In some embodiments, the cancer is retinoblastoma.

In some embodiments, the cancer is rhabdomyosarcoma.

In some embodiments, the cancer is salivary gland cancer.

In some embodiments, the cancer is skin cancer—basal and squamous cell.

In some embodiments, the cancer is skin cancer—melanoma.

In some embodiments, the cancer is small intestine cancer.

In some embodiments, the cancer is stomach (gastric) cancers.

In some embodiments, the cancer is testicular cancer.

In some embodiments, the cancer is thymus cancer.

In some embodiments, the cancer is thyroid cancer.

In some embodiments, the cancer is uterine sarcoma.

In some embodiments, the cancer is vaginal cancer.

In some embodiments, the cancer is vulvar cancer.

In some embodiments, the cancer is Wilms tumor.

In some embodiments, the cancer is laryngeal or hypopharyngeal cancer.

In some embodiments, the cancer is kidney cancer.

In some embodiments, the cancer is Kaposi sarcoma.

In some embodiments, the cancer is gestational trophoblastic disease.

In some embodiments, the cancer is gastrointestinal stromal tumor.

In some embodiments, the cancer is gastrointestinal carcinoid tumor.

In some embodiments, the cancer is gallbladder cancer.

In some embodiments, the cancer is eye cancer (melanoma and lymphoma).

In some embodiments, the cancer is Ewing tumor.

In some embodiments, the cancer is esophagus cancer.

In some embodiments, the cancer is endometrial cancer.

In some embodiments, the cancer is colorectal cancer.

In some embodiments, the cancer is cervical cancer.

In some embodiments, the cancer is brain or spinal cord tumor.

In some embodiments, the cancer is bone metastasis.

In some embodiments, the cancer is bone cancer.

In some embodiments, the cancer is bladder cancer.

In some embodiments, the cancer is bile duct cancer.

In some embodiments, the cancer is anal cancer.

In some embodiments, the cancer is adrenal cortical cancer.

In some embodiments, the disorder or disease is a neurologicalcondition/disorder/disease, wherein the neurologicalcondition/disorder/disease is selected from: Alzheimer's disease,frontotemporal dementias, dementia with Lewy bodies, prion diseases,Parkinson's disease, Huntington's disease, progressive supranuclearpalsy, corticobasal degeneration, multiple system atrophy, amyotrophiclateral sclerosis (ALS), inclusion body myositis, autism, degenerativemyopathies, diabetic neuropathy, other metabolic neuropathies, endocrineneuropathies, orthostatic hypotension, multiple sclerosis andCharcot-Marie-Tooth disease.

In some embodiments, the disorder or disease is a neurologicalcondition/disorder/disease associated with tau protein, amyloid,alpha-synuclein pathology, Tar DNA-binding Protein of 43KDa (TDP-43),Prion protein PrP or fused in sarcoma (FUS).

In some embodiments, the disorder or disease is a neurologicalcondition/disorder/disease, wherein the neurologicalcondition/disorder/disease is selected from the group consisting of:Alzheimer's Disease, Amyotrophic Lateral Sclerosis, Down Syndrome,Frontotemporal Dementia with Parkinsonism-17 (FTDP-17), Lewy bodydementia, Parkinson's Disease, Pick's Disease, and additional diseaseswith pronounced neurodegeneration such as Autism, Dementia, Epilepsy,Huntington's Disease, Multiple Sclerosis; diseases and disordersassociated with acquired brain injury such as Chronic TraumaticEncephalopathy, Traumatic Brain Injury, Tumor, and Stroke.

In some embodiments, the disorder or disease is a fibrotic disorder,wherein the fibrotic disorder is selected from the group consisting of:skin fibrosis; scleroderma; progressive systemic fibrosis; lungfibrosis; muscle fibrosis; kidney fibrosis; glomerulosclerosis;glomerulonephritis; hypertrophic scar formation; uterine fibrosis; renalfibrosis; cirrhosis of the liver, liver fibrosis; adhesions; chronicobstructive pulmonary disease; fibrosis following myocardial infarction;pulmonary fibrosis; fibrosis and scarring associated withdiffuse/interstitial lung disease; central nervous system fibrosis;fibrosis associated with proliferative vitreoretinopathy (PVR);restenosis; endometriosis; ischemic disease, and radiation fibrosis.

In some embodiments, the disorder or disease is chronic inflammationassociated with eye disorders, joint pain, arthritis (rheumatoid, osteo,psoriatic gout), cancers (colon, breast, lung, pancreas, and others),gastrointestinal disorders (ulcerative colitis and inflammatory boweldiseases), pulmonary disorders (chronic obstructive pulmonary disorderand asthma), allergies, skin disorders (atopic dermatitis andpsoriasis), diabetes, pancreatitis, tendonitis, hepatitis, heartdisease, myocarditis, stroke, lupus, and neurological disorders such asmultiple sclerosis, Parkinson's and dementia including Alzheimer'sdisease.

In some embodiments, a compound of Formula (I) inhibits DYRK1A.

In some embodiments, a compound of Formula (I) inhibits GSK3.

In some embodiments, a compound of Formula (I) inhibits GSK3β.

In some embodiments, a compound of Formula (I) inhibits DYRK1A andGSK3β.

In some embodiments, the compound of Formula (I) inhibits one or moreproteins in the Wnt pathway.

In some embodiments, the compound of Formula (I) inhibits signalinginduced by one or more Wnt proteins.

In some embodiments, the Wnt proteins are chosen from: WNT1, WNT2,WNT2B, WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A,WNT8B, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, and WNT16.

In some embodiments, the compound of Formula (I) inhibits a kinaseactivity.

In some embodiments, the method treats a disease or disorder mediated bythe Wnt pathway in a patient, the method comprises administering to thepatient a therapeutically effective amount of a compound (or compounds)of Formula (I), or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (I) inhibits one or moreWnt proteins.

In some embodiments, the method treats a disease or disorder mediated bykinase activity in a patient, the method comprises administering to thepatient a therapeutically effective amount of a compound (or compounds)of Formula (I), or a pharmaceutically acceptable salt thereof.

In some embodiments, the disease or disorder comprises tumor growth,cell proliferation, or angiogenesis.

In some embodiments, the method inhibits the activity of a proteinkinase receptor, the method comprises contacting the receptor with aneffective amount of a compound (or compounds) of Formula (I), or apharmaceutically acceptable salt thereof.

In some embodiments, the method treats a disease or disorder associatedwith aberrant cellular proliferation in a patient; the method comprisesadministering to the patient a therapeutically effective amount of acompound (or compounds) of Formula (I), or a pharmaceutically acceptablesalt thereof.

In some embodiments, the method prevents or reduces angiogenesis in apatient; the method comprises administering to the patient atherapeutically effective amount of a compound (or compounds) of Formula(I), or a pharmaceutically acceptable salt thereof.

In some embodiments, the method prevents or reduces abnormal cellularproliferation in a patient; the method comprises administering to thepatient a therapeutically effective amount of a compound (or compounds)of Formula (I), or a pharmaceutically acceptable salt thereof.

In some embodiments, the method treats a disease or disorder associatedwith aberrant cellular proliferation in a patient, the method comprisesadministering to the patient a pharmaceutical composition comprising oneor more of the compounds of claim 1 in combination with apharmaceutically acceptable carrier and one or more other agents.

Moreover, the compounds and compositions, for example, as inhibitors ofthe cyclin-dependent kinases (CDKs), can modulate the level of cellularRNA and DNA synthesis and therefore are expected to be useful in thetreatment of viral infections such as HIV, human papilloma virus, herpesvirus, Epstein-Barr virus, adenovirus, Sindbis virus, pox virus and thelike.

Compounds and compositions described herein can inhibit the kinaseactivity of, for example, CDK/cyclin complexes, such as those active inthe G₀ or G₁ stage of the cell cycle, e.g., CDK2, CDK4, and/or CDK6complexes.

Evaluation of Biological Activity

The biological activity of the compounds described herein can be testedusing any suitable assay known to those of skill in the art, see, e.g.,WO 2001/053268 and WO 2005/009997. For example, the activity of acompound may be tested using one or more of the test methods outlinedbelow.

In one example, tumor cells may be screened for Wnt independent growth.In such a method, tumor cells of interest are contacted with a compound(i.e. inhibitor) of interest, and the proliferation of the cells, e.g.by uptake of tritiated thymidine, is monitored. In some embodiments,tumor cells may be isolated from a candidate patient who has beenscreened for the presence of a cancer that is associated with a mutationin the Wnt signaling pathway. Candidate cancers include, withoutlimitation, those listed above.

In another example, one may utilize in vitro assays for Wnt biologicalactivity, e.g. stabilization of β-catenin and promoting growth of stemcells. Assays for biological activity of Wnt include stabilization ofβ-catenin, which can be measured, for example, by serial dilutions of acandidate inhibitor composition. An exemplary assay for Wnt biologicalactivity contacts a candidate inhibitor with cells containingconstitutively active Wnt/β-catenin signaling. The cells are culturedfor a period of time sufficient to stabilize β-catenin, usually at leastabout 1 hour, and lysed. The cell lysate is resolved by SDS PAGE, thentransferred to nitrocellulose and probed with antibodies specific forβ-catenin.

In a further example, the activity of a candidate compound can bemeasured in a Xenopus secondary axis bioassay (Leyns, L. et al. Cell(1997), 88(6), 747-756).

In another example, in vitro assays for DYRK1A biological activity maybe used, e.g. regulation of microtubule-associated protein tau(MAPT/Tau) phosphorylation in neuronal cell line such as the humanSH-SY5Y neuroblastoma cell line. Assays for DYRK1A-regulated level ofphosphorylation can include monitoring levels of basal pSer396 Tau,which can be measured, for example, by serial dilutions of a candidateinhibitor composition using a ten micromolar top concentration anddetected by ELISA or Western Blotting. An exemplary assay forDYRK-1A-regulated phosphorylation uses the SH-SY5Y cells cultured in a96 well plate format for a period of time sufficient to stabilizemicrotubules and Tau phosphorylation, usually at least 2 days, thentreated with a ⅓ serial dilution of compounds overnight and lysed. Thecell lysate is resolved by SDS PAGE, then transferred to nitrocelluloseand probed with an antibody specific for pSer396 Tau. Thechemiluminescence signal for HRP-linked antibodies used in westernblotting is detected using a Carestream Image Station and blotdensitometry for pSer396 and beta-actin are analyzed using ImageJ (NIH).

In a further example, the activity of a candidate compound can bemeasured by ELISA by adding the lysate mentioned above onto totalTau-coated plates and detected with a specific pSer396 antibody.Colorimetric detection of ELISA signal is performed by Cytation3 platereader (Biotek).

To further illustrate this disclosure, the following examples areincluded. The examples should not, of course, be construed asspecifically limiting the disclosure. Variations of these exampleswithin the scope of the claims are within the purview of one skilled inthe art and are considered to fall within the scope of the disclosure asdescribed, and claimed herein. The reader will recognize that theskilled artisan, armed with the present disclosure, and skill in the artis able to prepare and use the disclosure without exhaustive examples.

EXAMPLES

Compound Preparation

The starting materials used in preparing the compounds of the disclosureare known, made by known methods, or are commercially available. It willbe apparent to the skilled artisan that methods for preparing precursorsand functionality related to the compounds claimed herein are generallydescribed in the literature. The skilled artisan given the literatureand this disclosure is well equipped to prepare any of the compounds.

It is recognized that the skilled artisan in the art of organicchemistry can readily carry out manipulations without further direction,that is, it is well within the scope and practice of the skilled artisanto carry out these manipulations. These include reduction of carbonylcompounds to their corresponding alcohols, oxidations, acylations,aromatic substitutions, both electrophilic and nucleophilic,etherifications, esterification and saponification and the like. Thesemanipulations are discussed in standard texts such as March's AdvancedOrganic Chemistry: Reactions, Mechanisms, and Structure 7^(th) Ed., JohnWiley & Sons (2013), Carey and Sundberg, Advanced Organic Chemistry5^(th) Ed., Springer (2007), Comprehensive Organic Transformations: AGuide to Functional Group Transformations, 2^(nd) Ed., John Wiley & Sons(1999) (incorporated herein by reference in its entirety) and the like.

The skilled artisan will readily appreciate that certain reactions arebest carried out when other functionality is masked or protected in themolecule, thus avoiding any undesirable side reactions and/or increasingthe yield of the reaction. Often the skilled artisan utilizes protectinggroups to accomplish such increased yields or to avoid the undesiredreactions. These reactions are found in the literature and are also wellwithin the scope of the skilled artisan. Examples of many of thesemanipulations can be found for example in P. Wuts Greene's ProtectiveGroups in Organic Synthesis, 5th Ed., John Wiley & Sons (2014),incorporated herein by reference in its entirety.

Trademarks used herein are examples only and reflect illustrativematerials used at the time of the disclosure. The skilled artisan willrecognize that variations in lot, manufacturing processes, and the like,are expected. Hence the examples, and the trademarks used in them arenon-limiting, and they are not intended to be limiting, but are merelyan illustration of how a skilled artisan may choose to perform one ormore of the embodiments of the disclosure.

(¹H) nuclear magnetic resonance spectra (NMR) were measured in theindicated solvents on a Bruker NMR spectrometer (Avance™ DRX300, 300 MHzfor ¹H or Avance™ DRX500, 500 MHz for ¹H) or Varian NMR spectrometer(Mercury 400BB, 400 MHz for ¹H). Peak positions are expressed in partsper million (ppm) downfield from tetramethylsilane. The peakmultiplicities are denoted as follows, s, singlet; d, doublet; t,triplet; q, quartet; ABq, AB quartet; quin, quintet; sex, sextet; sep,septet; non, nonet; dd, doublet of doublets; ddd, doublet of doublets ofdoublets; d/ABq, doublet of AB quartet; dt, doublet of triplets; td,triplet of doublets; dq, doublet of quartets; m, multiplet.

The following abbreviations have the indicated meanings:

-   -   brine=saturated aqueous sodium chloride    -   CDCl₃=deuterated chloroform    -   DCE=dichloroethane    -   DCM=dichloromethane    -   DIPEA=N,N-diisopropylethylamine    -   DMA=dimethylacetamide    -   DMAP=4-dimethylaminopyridine    -   DMF=N,N-dimethylformamide    -   DMSO-d₆=deuterated dimethylsulfoxide    -   ESIMS=electron spray mass spectrometry    -   EtOAc=ethyl acetate    -   HATU=1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium        3-oxid hexafluorophosphate    -   HCl=hydrochloric acid    -   HOAc=acetic acid    -   ISCO=Teledyne ISCO, Inc brand CombiFlash® Rf 200    -   KOAc=potassium acetate    -   LC/MS=Liquid chromatography-mass spectrometry    -   MeCN=acetonitrile    -   MeOH=methanol    -   MW=microwave irradiation    -   NaBH₃CN=sodium cyanoborohydride    -   NaHCO₃=sodium bicarbonate    -   NMR=nuclear magnetic resonance    -   ON=overnight    -   Pd(dppf)Cl₂=1,1′-bis(diphenylphosphino)ferrocene-palladium(II)dichloride    -   Pd(PPh₃)₄=tetrakis(triphenylphosphine)palladium(0)    -   r.t.=room temperature    -   TEA=triethylamine    -   THF=tetrahydrofuran    -   TLC=thin layer chromatography

The following example schemes are provided for the guidance of thereader, and collectively represent an example method for making thecompounds provided herein. Furthermore, other methods for preparingcompounds of the disclosure will be readily apparent to the person ofordinary skill in the art in light of the following reaction schemes andexamples. The skilled artisan is thoroughly equipped to prepare thesecompounds by those methods given the literature and this disclosure. Thecompound numberings used in the synthetic schemes depicted below aremeant for those specific schemes only, and should not be construed as orconfused with same numberings in other sections of the application.Unless otherwise indicated, all variables are as defined above.

General Procedures

Compounds of Formula I of the present disclosure can be prepared asdepicted in Scheme 1.

Scheme 1 describes a method for preparation ofisoquinoline-3-carboxamide derivatives (IX) by first coupling the aminewith a variety of acids (III) to produce amide IV. The bromo derivativeIV is then reacted with bis(pinacolato)diboron to give the pinacol ester(V). Suzuki coupling with a variety of 5-membered heteroaryl bromides(VIII) yields the desired R³ substituted isoquinoline IX. Alternatively,the bromo derivative IV is Suzuki coupled with a variety of 5-memberedheteroaryl pinacol esters (VI) or coupled to a variety of 5-memberedheteroaryl stannanes (VII) to produce the final R³ substitutedisoquinoline IX.

In other embodiments, compounds of Formula I of the present disclosurecan be prepared as depicted in Scheme 2.

Scheme 2 describes a method for preparation ofisoquinoline-3-carboxamide derivatives (IXa) starting with bromointermediate IV or IVa and couple with the nitrogen of a variety of R³NH heteroaryls to produce the final R³ substituted isoquinoline IXa.

ILLUSTRATIVE COMPOUND EXAMPLES

Preparation of intermediate 6-bromoisoquinolin-1-d-3-amine (XIV) isdepicted below in Scheme 3.

Step 1

To a mixture of 1,6-dibromoisoquinolin-3-amine (XIII) (0.5 g, 1.66mmol), ammonium formate-d₅ (0.56 g, 8.28 mmol) and Pd(PPh₃)₄ (191.3 mg,0.170 mmol) in DMF (5 mL) was heated to 50° C. for 48 h. The solventswere concentrated and the residue was suspended in chloroform. The solidwas collected by filtration and washed with water and EtOAc. The solidwere dried under high vacuo to obtain6-bromo-1-deuterio-isoquinolin-3-amine (XIV) (115 mg, 0.513 mmol, 31.0%yield) as a pale yellow solid. ¹H NMR (500 MHz, DMSO-d₆) δ ppm 6.11 (2H,s), 6.55 (1H, s), 7.22 (1H, dd, J=8.78, 1.92 Hz), 7.73 (1H, d, J=8.51Hz), 7.79 (1H, d, J=1.92 Hz); ESIMS found for C₉H₆DBrN₂ m/z 224.0(⁷⁹BrM+H).

Preparation of intermediate 6-bromo-4-chloroisoquinolin-3-amine (XVI) isdepicted below in Scheme 4.

Step 1

To a stirred suspension of 6-bromoisoquinolin-3-amine (XV) (1.0 g, 4.48mmol) in DMF (15 mL) at 0° C. was added 1-chloropyrrolidine-2,5-dione(598.6 mg, 4.48 mmol) portionwise. The mixture was stirred at 0° C. for6 h. The reaction mixture was added to water (150 mL), stirred for 1 hand the resulting solids were collected by filtration and air driedovernight to obtain 6-bromo-4-chloro-isoquinolin-3-amine (XVI) (922 mg,3.58 mmol, 79.9% yield) as a beige solid which was used for next stepwithout purification. ¹H NMR (499 MHz, DMSO-d₆) δ ppm 6.55 (2H, s), 7.40(1H, dd, J=8.64, 1.78 Hz), 7.88 (1H, d, J=8.51 Hz), 7.90 (1H, d, J=1.10Hz), 8.86 (1H, s); ESIMS found for C₉H₆BrClN₂ m/z 256.9 (⁷⁹BrM+H).

Preparation of intermediate 6-bromo-4-methylisoquinolin-3-amine (XVIII)is depicted below in Scheme 5.

Step 1

To a stirred suspension of 6-bromoisoquinolin-3-amine (XV) (2.g, 8.97mmol) in DMF (25.1 mL) at 0° C. was added 1-iodopyrrolidine-2,5-dione(2.02 g, 8.97 mmol) portionwise, The mixture was stirred at 0° C. for 1hr. LC-MS of the mixture showed completion of the reaction and thedesired product. The solvent was removed under vacuum, the residue waspurified by C18 Silica gel (240 g) [0-100% H₂O/MeCN (0.1% Formic acid)]to produce 6-bromo-4-iodo-isoquinolin-3-amine (XVII) (1.95 g, 5.58 mmol,62.2% yield) as a brown solid. ¹H NMR (499 MHz, DMSO-d₆) δ ppm 6.41 (2H,br s), 7.40 (1H, dd, J=8.64, 1.78 Hz), 7.76-7.81 (1H, m), 7.82 (1H, d,J=8.51 Hz), 8.81 (1H, s); ESIMS found for C₉H₆BrIN₂ m/z 348.9 (⁷⁹BrM+H).

Step 2

A stirred solution of 6-bromo-4-iodo-isoquinolin-3-amine (XVII) (1.0 g,2.87 mmol), 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (0.72 g, 2.87mmol), Pd(dppf)Cl₂ (0.23 g, 0.29 mmol), and K₃PO₄ (5.73 mL, 5.73 mmol)in 1,4-dioxane (10 mL) was heated to 90° C. for 3 days. The solvent wasremoved under high vacuum and the residue was purified by C18 silica gel(240 g) [0→20% H₂O/MeCN (0.1% Formic acid)] to produce6-bromo-4-methyl-isoquinolin-3-amine (XVIII) (74 mg, 0.312 mmol, 10.9%yield) as an off-white solid. ¹H NMR (499 MHz, DMSO-d₆) δ ppm 2.23 (3H,br s), 5.91 (2H, br s), 7.27 (1H, br d, J=2.20 Hz), 7.71-7.82 (1H, m),7.92 (1H, br s), 8.72 (1H, br s); ESIMS found for C₁₀H₉BrN₂ m/z 239.0(^(s8)BrM+H).

Preparation of intermediate 6-bromo-7-fluoroisoquinoline-3-amine (XXI)is depicted below in Scheme 6.

Step 1

To a vial was added 2,2-diethoxyacetonitrile (XIX) (1.0 g, 7.74 mmol)dissolved MeOH (7.74 mL) followed by addition of MeONa/MeOH (0.18 mL,0.77 mmol) dropwise. The reaction was stirred at room temperature for 20h. HOAc (44.3 μL, 0.77 mmol) was added until pH=7-8 (using pH strips).(4-Bromo-3-fluoro-phenyl)methanamine hydrochloride (XX) (1.86 g, 7.74mmol) was added and stirred at 40° C. for 4 h. The solvent was removedunder vacuum. Sulfuric acid (12.6 mL, 232.3 mmol) was added and stirredat 40° C. for 16 h. NH₄OH (30.8 mL, 240.0 mmol) was added dropwise at 0°C. The solvent was removed under vacuum and the residue was purified byC18 silica gel (240 g) [0→50% H₂O/MeCN (0.1% Formic acid)] to produce6-bromo-7-fluoro-isoquinolin-3-amine (XXI) (1.33 g, 5.50 mmol, 71.1%yield) as an off-white solid. ¹H NMR (499 MHz, DMSO-d₆) δ ppm 6.07 (2H,s), 6.61 (1H, s), 7.76 (1H, d, J=9.33 Hz), 8.01 (1H, d, J=6.86 Hz), 8.80(1H, s); ESIMS found for C₉H₆BrFN₂ m/z 242.9 (⁸¹BrM+H).

Preparation of intermediates 6-bromo-7-chloroisoquinolin-3-amine (XXIII)and 6-bromo-5-chloroisoquinolin-3-amine (XXIV) is depicted below inScheme 7.

Step 1

To a stirred solution of 2,2-diethoxyacetonitrile (XIX) (0.59 g, 4.57mmol) in a vial containing MeOH (4.57 mL) was added MeONa (0.1 mL, 0.46mmol) dropwise. The reaction was stirred at 35° C. for 20 h. HOAc wasadded (26.1 μL, 0.46 mmol) (checked that the pH is 7-8 using pH strips)followed by (4-bromo-3-chloro-phenyl)methanamine (XXII) (1.01 g, 4.57mmol). The mixture was stirred at 35° C. for 40 h. The solvent wasremoved under vacuum. Sulfuric Acid (7.43 mL, 137.0 mmol) was then addedand stirred at 35° C. for 16 h. NH₄OH (60.6 mL, 141.6 mmol) was added at0° C. The reaction was filtered through Celite and purified by C18silica gel (240 g) [0→30% H₂O/MeCN (0.1% Formic acid)] to produce a 1:1mixture (by NMR) of 6-bromo-7-chloro-isoquinolin-3-amine (XXIII) and6-bromo-5-chloroisoquinolin-3-amine (XXIV) (633.7 mg, 2.46 mmol, 53.9%yield). ¹H NMR (499 MHz, DMSO-d₆) δ ppm 6.23 (2H, s), 6.46 (2H, s), 6.57(1H, s), 6.83 (1H, s), 7.40 (1H, d, J=8.51 Hz), 7.74 (1H, d, J=8.51 Hz),8.05 (1H, s), 8.09 (1H, s), 8.81 (1H, s), 8.88 (1H, s); ESIMS found forC₉H₆BrClN₂ m/z 256.9 (⁷⁹BrM+H).

Preparation of intermediates 6-bromo-7-methylisoquinolin-3-amine (XXVI)and 6-bromo-5-methylisoquinolin-3-amine (XXVII) is depicted below inScheme 8.

Step 1

To a stirred solution of 2,2-diethoxyacetonitrile (XIX) (0.33 g, 2.52mmol) in a vial containing MeOH (2.52 mL) was added MeONa (0.23 mL, 0.25mmol) dropwise. The reaction was stirred at 22° C. for 20 h. HOAc wasadded (14.4 μL, 0.25 mmol) (checked that the pH is 7-8 using pH strips)followed by (4-bromo-3-methyl-phenyl)methanamine (XXV) (0.5 g, 2.52mmol). The mixture was stirred at 40° C. for 40 h. The solvent wasremoved under vacuum. Sulfuric Acid (4.09 mL, 75.49 mmol) was then addedand stirred at 40° C. for 16 h. NH₄OH (33.4 mL, 78 mmol) was added at 0°C. The reaction was filtered through Celite and purified by C18 silicagel (240 g) [0→30% H₂O/MeCN (0.1% Formic acid)] to produce a 1:1 mixture(by NMR) of 6-bromo-7-methylisoquinolin-3-amine (XXVI) and6-bromo-5-methylisoquinolin-3-amine (XXVII) (378 mg, 1.59 mmol, 63.4%yield). ¹H NMR (499 MHz, DMSO-d₆) δ ppm 2.40 (3H, s), 2.52 (3H, s), 5.96(2H, s), 6.12 (1H, s), 6.54 (1H, s), 6.71 (1H, s), 7.27 (1H, d, J=8.78Hz), 7.58 (1H, d, J=8.78 Hz), 7.73 (1H, s), 7.86 (1H, s), 8.74 (1H, s),8.79 (1H, s); ESIMS found for C₁₀H₉BrN₂ m/z 237.0 (⁷⁹BrM+H).

Preparation of intermediate1-(1-(tert-butoxycarbonyl)piperidin-4-yl)-1H-pyrazole-4-carboxylic acid(XXXI) is depicted below in Scheme 9.

Step 1

To a suspension of NaH (0.24 g, 5.99 mmol) in DMF (15 mL) at 0° C. underargon was added ethyl 1H-pyrazole-4-carboxylate (XXXVIII) (0.7 g, 5mmol). After stirring for 30 min, tert-butyl4-(p-tolylsulfonyloxy)piperidine-1-carboxylate (XXIX) (2.13 g, 5.99mmol) was added and the mixture heated at 80° C. for 1 h. The reactionwas poured into water and extracted with EtOAc. The organic layer waswashed with brine, dried, filtered and concentrated under vacuum. Thecrude product was purified by silica gel chromatography (0-40%EtOAc/Hexanes) to afford tert-butyl4-(4-(ethoxycarbonyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (XXX) asa white solid (1.25 g, 3.87 mmol, 77.4% yield). ESIMS found forC₁₆H₂₅N₃O₄ m/z 346.2 (M+Na).

Step 2

To a solution of tert-butyl4-(4-(ethoxycarbonyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (XXX)(1.25 g, 3.87 mmol) in MeOH (4 mL) and THF (4 mL) was added LiOH (3.87mL, 15.46 mmol). The reaction stirred at 60° C. for 2 h. The mixture wasconcentrated and the residue triturated in water. The solution wasacidified with 2 N HCl and the resulting solid was filtered to afford1-(1-(tert-butoxycarbonyl)piperidin-4-yl)-1H-pyrazole-4-carboxylic acid(XXXI) as a white solid (1.04 g, 3.52 mmol, 91.1% yield). ESIMS foundfor C₁₄H₂₁N₃O₄ m/z 318.1 (M+Na).

Preparation of intermediate2-(1-(tert-butoxycarbonyl)piperidin-4-yl)oxazole-4-carboxylic acid(XXXIV) is depicted below in Scheme 10.

Step 1

To a suspension of ethyl 2-(piperidin-4-yl)oxazole-4-carboxylate (XXXII)(purchased from CombiBlocks) (1.0 g, 4.46 mmol) and di-tert-butyldicarbonate (1.27 g, 5.8 mmol) in THF (15 mL) was added TEA (0.93 mL,6.69 mmol). The mixture was stirred at 65° C. for 16 h before themixture was poured into saturated NaHCO₃. The mixture was extracted withEtOAc (2×) and the combined organic layers were washed with brine,dried, filtered and concentrated under vacuum. The crude product waspurified by silica gel chromatography (0-50% EtOAc/Hexanes) to affordethyl 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)oxazole-4-carboxylate(XXXIII) as a colorless oil (solidified to a white solid while standingat room temperature) (1.4 g, 4.32 mmol, 96.8% yield). ESIMS found forC₁₆H₂₄N₂O₅ m/z 346.95 (M+Na).

Step 2

To a solution of ethyl2-(1-(tert-butoxycarbonyl)piperidin-4-yl)oxazole-4-carboxylate (XXXIII)(1.41 g, 4.35 mmol) in MeOH (6 mL) and THF (6 mL) was added LiOH (4.35mL, 17.39 mmol) and the mixture stirred at 60° C. for 1 h. The mixturewas concentrated to remove the organic solvents. The residual water wasacidified with 2 N HCl and the resulting solid was filtered to afford2-(1-(tert-butoxycarbonyl)piperidin-4-yl)oxazole-4-carboxylic acid(XXXIV) as a white solid (1.1 g, 3.71 mmol, 85.4% yield). ESIMS foundfor C₁₄H₂₀N₂O₅ m/z 319.0 (M+Na).

Preparation of intermediate2-(3-(dimethylamino)azetidin-1-yl)thiazole-4-carboxylic acid (XXXIX) isdepicted below in Scheme 11.

Step 1

To a solution of 2-chlorothiazole-5-carboxylic acid (XXXV) (1.0 g, 6.11mmol), di-tert-butyl dicarbonate (3.07 g, 14.06 mmol) in THF (19.8 mL)was added DMAP (0.15 g, 1.22 mmol). The reaction was heated at 60° C.for 1 h. The solvent was removed under vacuum and the residue waspurified by silica gel (40 g) (0-50% EtOAc/hexanes) to producetert-butyl 2-chlorothiazole-4-carboxylate (XXXVI) as a clear liquid(1.05 g, 4.79 mmol, 78.4% yield). ESIMS found for C₈H₁₀ClNO₂S m/z 220.0(M+H).

Step 2

To a suspension of tert-butyl 2-chlorothiazole-4-carboxylate (XXXVI)(265 mg, 1.21 mmol) in DMSO (10 mL) was addedN,N-dimethylazetidin-3-amine (XXXVII) (313.2 mg, 1.81 mmol) and DIPEA(1.05 mL, 6.03 mmol). The mixture was heated at reflux for 1 day and themixture was cooled to room temperature. The reaction was poured intowater and the aqueous layer was extracted with EtOAc. The organic layerwas dried and evaporated under vacuum. The residue was purified bycolumn chromatography (0-40% 20% MeOH (7 N NH₃)—CHCl₃/CHCl₃) to affordtert-butyl 2-(3-(dimethylamino)azetidin-1-yl)thiazole-4-carboxylate(XXXVIII) as a brown oil (67 mg, 0.24 mmol, 19.6% yield). ESIMS foundfor C₁₃H₂₁N₃O₂S m/z 284.1 (M+H).

Step 3

To a stirred solution of tert-butyl2-(3-(dimethylamino)azetidin-1-yl)thiazole-4-carboxylate (XXXVIII) (0.2g, 0.71 mmol) in dioxane (15 mL) was added HCl (4 N in dioxane) (1.76mL, 7.06 mmol). The reaction was heated at 100° C. for 16 h. The whitesolid was filtered and washed with MTBE, brine and dried to produce2-(3-(dimethylamino)azetidin-1-yl)thiazole-4-carboxylic acid (XXXIX) asa white solid (0.45 g, 1.50 mmol, 212% yield). ESIMS found forC₉H₁₃N₃O₂S m/z 228.1 (M+H).

The following intermediates were prepared in accordance with theprocedure described in the above Scheme 11.

2-(3-(Dimethylamino)azetidin-1-yl)oxazole-4-carboxylic acid (XL): Whitesolid, (543.9 mg, 1.91 mmol, 90.1% yield). ESIMS found C₉H₁₃N₃O₃ m/z212.0 (M+H).

Preparation of intermediate1-(methyl-d3)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(XLII) is depicted below in Scheme 12.

Step 1

To a stirred suspension of4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (XLI) (1.435g, 7.4 mmol) and Cs₂CO₃ (2.89 g, 8.87 mmol) in DMF (15 mL) was addedtrideuterio(iodo)methane (0.51 mL, 8.13 mmol) and the mixture wasstirred at room temperature overnight. The reaction mixture was filteredand the filtrates were concentrated and dried under high vacuo to obtain4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(trideuteriomethyl)pyrazole(XLII) (3.9 g, 18.48 mmol, 249.8% yield) as a white solid which was usedfor next step without purification. ESIMS found for C₁₀H₁₄[²H₃]BN₂O₂ m/z212. (M+1).

Example 1

Preparation ofN-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(piperidin-4-yl)-1H-pyrazole-4-carboxamide(2),1-(1-isopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide(8),1-(1-cyclopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide(9), and isopropyl4-(4-((6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)carbamoyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate(10) is depicted below in Scheme 13.

Step 1

In a sealed tube, 6-bromoisoquinolin-3-amine (XV) (10.0 g, 45.01 mmol)was added to1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(XLIII) (10.58 g, 50.85 mmol), 12 M aqueous solution of K₃PO₄ (9.52 mL,114.95 mmol), Pd(dppf)Cl₂ (1.95 g, 2.39 mmol) in dioxane (240 mL) andwater (59.4 mL). This mixture was degassed with Argon, sealed and heatedto 90° C. for 16 h. The mixture was poured into water (500 mL), thesolid was filtered and rinsed with MeOH to yield6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-amine (XLIV) (5.69 g, 25.37mmol, 56.4% yield). The product was used for the next step withoutfurther purification. ESIMS found for C₁₃H₁₂N₄ m/z 225.0 (M+H).

Step 2

To a suspension of 6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-amine(XLIV) (0.4 g, 1.78 mmol),1-(1-(tert-butoxycarbonyl)piperidin-4-yl)-1H-pyrazole-4-carboxylic acid(XXXI) (0.58 g, 1.96 mmol), DMAP (0.22 g, 1.78 mmol) and HATU (0.81 g,2.14 mmol) in DMF (9 mL) was added DIPEA (1.24 mL, 7.13 mmol). Theresulting mixture was stirred 80° C. for 3 h. An additional 1.2 equiv.of HATU was added and the reaction mixture stirred at 80° C. for another16 h. The mixture was poured into water and the solid precipitatefiltered. The filtrate was extracted with EtOAc (2×) and the combinedorganic layers were washed with brine, dried, filtered and concentratedunder vacuum. The crude product was purified by silica gelchromatography (0→100% EtOAc/Hexanes) to afford tert-butyl4-[4-[[6-(1-methylpyrazol-4-yl)-3-isoquinolyl]carbamoyl]pyrazol-1-yl]piperidine-1-carboxylate(XLV) as a reddish brown solid (0.70 g, 1.40 mmol, 78.2% yield). ESIMSfound for C₂₇H₃₁N₇O₃ m/z 502.25 (M+H).

Step 3

To a suspension of tert-butyl4-[4-[[6-(1-methylpyrazol-4-yl)-3-isoquinolyl]carbamoyl]pyrazol-1-yl]piperidine-1-carboxylate(XLV) (700 mg, 1.4 mmol) in MeOH (10 mL) was added HCl (4 M in dioxane)(4.54 mL, 18.14 mmol). The resulting solution was stirred at 60° C. for2 h and then concentrated under vacuum. The HCl salt was neutralizedwith 7 N NH₃/MeOH and the crude product absorbed onto Celite andpurified by silica gel chromatography (0→15% 7 N NH₃-MeOH/CHCl₃). Thefractions containing the product were concentrated and residuetriturated in ether. The resulting solid was filtered to affordN-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(piperidin-4-yl)-1H-pyrazole-4-carboxamide2 as a white solid (370.0 mg, 0.876 mmol, 62.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.79 (2H, qd, J=11.98, 3.84 Hz), 2.01 (2H, br dd,J=11.80, 2.20 Hz), 2.63 (2H, td, J=12.28, 2.06 Hz), 3.04-3.09 (2H, m),3.91 (3H, s), 4.22-4.31 (1H, m), 7.77 (1H, dd, J=8.51, 1.65 Hz), 8.03(1H, d, J=8.78 Hz), 8.08 (1H, s), 8.10 (1H, s), 8.17 (1H, s), 8.37 (1H,s), 8.54 (1H, s), 8.59 (1H, s), 9.08 (1H, s), 10.49 (1H, s); ESIMS foundfor C₂₂H₂₃N₇O m/z 402.2 (M+1).

Step 4

To a suspension ofN-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(piperidin-4-yl)-1H-pyrazole-4-carboxamide2 (148 mg, 0.37 mmol) and Cs₂CO₃ (180.2 mg, 0.55 mmol) in DMF (3 mL) wasadded 2-iodopropane (0.05 mL, 0.48 mmol). The mixture was stirred at 65°C. for 2 h and the mixture diluted with water and extracted with EtOAc(2×). The combined organic layers were washed with brine, dried,filtered and concentrated. The crude product was purified by silica gelchromatography (0-6% 7 N NH₃-MeOH/CHCl₃). The fractions containing theproducts were concentrated and the residue triturated in ether to afford1-(1-isopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide8 as a yellow solid (75.0 mg, 0.169 mmol, 45.9% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 0.99 (6H, d, J=6.59 Hz), 1.89 (2H, qd, J=11.89, 3.84 Hz),2.03-2.10 (2H, m), 2.24-2.31 (2H, m), 2.75 (1H, spt, J=6.54 Hz), 2.88(2H, br d, J=11.80 Hz), 3.91 (3H, s), 4.15 (1H, tt, J=11.42, 4.22 Hz),7.77 (1H, dd, J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.51 Hz), 8.08 (1H, s),8.10 (1H, s), 8.17 (1H, s), 8.37 (1H, s), 8.54 (1H, s), 8.60 (1H, s),9.08 (1H, s), 10.47 (1H, s); ESIMS found for C₂₅H₂₉N₇O m/z 444.2 (M+1);and isopropyl4-(4-((6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)carbamoyl)-1H-pyrazol-1-yl)piperidine-1-carboxylate10 as a white solid (20.0 mg, 0.041 mmol, 11.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.21 (6H, d, J=6.04 Hz), 1.80 (2H, qd, J=12.08, 4.39 Hz),2.08 (2H, br dd, J=12.35, 2.20 Hz), 2.99 (2H, br s), 3.91 (3H, s),4.03-4.13 (2H, m), 4.44 (1H, tt, J=11.29, 3.95 Hz), 4.80 (1H, spt,J=6.22 Hz), 7.77 (1H, dd, J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.51 Hz),8.08 (1H, s), 8.10 (1H, s), 8.19 (1H, s), 8.37 (1H, s), 8.53 (1H, s),8.62 (1H, s), 9.08 (1H, s), 10.48 (1H, s); ESIMS found for C₂₆H₂₉N₇O₃m/z 488.2 (M+1).

Step 5

A solution ofN-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(piperidin-4-yl)-1H-pyrazole-4-carboxamide2 (100 mg, 0.25 mmol), (1-ethoxycyclopropoxy)trimethylsilane (XLVI)(0.25 mL, 1.25 mmol) and HOAc (0.04 mL, 0.75 mmol) in MeOH (4 mL) wasstirred at room temperature for 10 min. To the mixture was added withNaCNBH₃ (78.2 mg, 1.25 mmol) and the mixture was stirred at 60° C. for 3h. The mixture was cooled to room temperature and concentrated undervacuum. The residue was purified by chromatography (0-10% 7NNH₃-MeOH/CHCl₃). The fractions containing the product were concentrated,the residue triturated in ether and the solid filtered and dried to give1-(1-cyclopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide9 as a white solid (32.0 mg, 0.069 mmol, 27.6% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 0.29-0.36 (2H, m), 0.40-0.48 (2H, m), 1.66 (1H, tt,J=6.62, 3.53 Hz), 1.87 (2H, qd, J=11.98, 3.84 Hz), 2.01-2.08 (2H, m),2.35 (2H, td, J=11.80, 1.92 Hz), 3.02 (2H, br d, J=11.80 Hz), 3.91 (3H,s), 4.21 (1H, tt, J=11.32, 4.05 Hz), 7.76 (1H, dd, J=8.51, 1.65 Hz),8.03 (1H, d, J=8.78 Hz), 8.07 (1H, s), 8.10 (1H, s), 8.17 (1H, s), 8.37(1H, s), 8.53 (1H, s), 8.59 (1H, s), 9.08 (1H, s), 10.46 (1H, s); ESIMSfound for C₂₅H₂₇N₇O m/z 442.0 (M+1).

Example 2

Preparation ofN-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)acetamide(47) is depicted below in Scheme 14.

Step 1

A mixture of 6-bromoisoquinolin-3-amine (XV) (2.57 g, 11.52 mmol),1-methyl-5-(tributylstannyl)-1H-imidazole (XLVII) (Purchased fromCombi-blocks) (4.28 g, 11.53 mmol), Pd(PPh₃)₄ (1.39 g, 1.2 mmol) indioxane (57.6 mL) was purged with N₂ gas for 10 min and then heated to90° C. overnight. The solvent was evaporated under high vacuum and theresidue was purified by column chromatography (0-10% MeOH/CHCl₃) to give6-(3-methylimidazol-4-yl)isoquinolin-3-amine (XLVIII) as an olive greensolid (2.32 g, 9.83 mmol, 85.3% yield). ESIMS found for C₁₃H₁₂N₄ m/z225.1 (M+H).

Step 2

To a suspension of 6-(3-methylimidazol-4-yl)isoquinolin-3-amine (XLVIII)(100 mg, 0.45 mmol), 2-(1-methylpyrazol-4-yl)acetic acid (XLIX(purchased from Enamine) (75 mg, 0.54 mmol), DMAP (27.2 mg, 0.22 mmol)and HATU (203.5 mg, 0.54 mmol) in DMF (5 mL) was added DIPEA (0.19 mL,1.1 lmmol). The resulting mixture was stirred at 80° C. for 1.5 h. Thereaction mixture was cooled to room temperature and poured into water.The resulting solid was filtered and purified by silica gelchromatography (0-10% MeOH/CHCl₃). The fractions containing the productwere concentrated. The product was further purified by HPLC (0→35%MeCN/water) and the fractions containing the product concentrated,re-dissolved in MeOH and filtered through bicarbonate resin. Thefiltrate was concentrated and triturated in ether, filtered and driedunder high vacuo to affordN-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)acetamide47 as a white solid (20.0 mg, 0.058 mmol, 12.9% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 3.59 (2H, s), 3.79 (3H, s), 3.82 (3H, s), 7.30 (1H, d,J=0.82 Hz), 7.36 (1H, s), 7.61 (1H, s), 7.67 (1H, dd, J=8.51, 1.65 Hz),7.79 (1H, s), 7.99 (1H, s), 8.09 (1H, d, J=8.51 Hz), 8.50 (1H, s), 9.13(1H, s), 10.66 (1H, s); ESIMS found for C₁₉H₁₈N₆O m/z 347.2 (M+1).

Example 3

Preparation of2-(1H-imidazol-1-yl)-N-(6-(2-methyloxazol-5-yl)isoquinolin-3-yl)acetamide(64) is depicted below in Scheme 15.

Step 1

To a suspension of 6-bromoisoquinolin-3-amine (XV) (10 g, 44.83 mmol)and 2-chloroacetic anhydride (L) (9.2 g, 53.79 mmol) in dry THF (100 mL)was added TEA (15.62 mL, 112.07 mmol). The mixture was stirred at roomtemperature for 16 h then poured into a mixture of aqueous saturatedNaHCO₃ (200 mL) and water (300 mL). The mixture was stirred for an hourand the resulting solids were collected by filtration, air-dried andsuspended in diethyl ether (400 mL), stirred for 30 min. The solidfiltered and dried under high vacuo to obtainN-(6-bromoisoquinolin-3-yl)-2-chloroacetamide (LI) as a brown solid(9.36 g, 31.25 mmol, 69.7% yield). ESIMS found for C₁₁H₈BrClN₂O m/z298.95 (M+1).

Step 2

A mixture of N-(6-bromoisoquinolin-3-yl)-2-chloroacetamide (LI) (0.15 g,0.481 mmol, 96.0% yield), imidazole (LII) (0.1 mL, 1.5 mmol), and DIPEA(0.17 mL, 1 mmol) in MeCN (2 mL) was heated at 110° C. for 30 min undermicrowave irradiation. The reaction concentrated under vacuum to giveN-(6-bromoisoquinolin-3-yl)-2-(1H-imidazol-1-yl)acetamide (LIII) (165.3mg, 0.50 mmol). The reaction mixture was used for next reaction withoutfurther purification. ESIMS found for C₁₄H_(1l)BrN₄O m/z 331.0 (M+H).

Step 3

To a mixture ofN-(6-bromoisoquinolin-3-yl)-2-(1H-imidazol-1-yl)acetamide (LIII) (172mg, 0.50 mmol),2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)oxazole (LIV)(130.7 mg, 0.63 mmol), and Pd(dppf)Cl₂ (40.8 mg, 0.05 mmol) in MeCN (1mL) was added 2 M aqueous solution of K₂CO₃ (0.63 mL, 1.25 mmol). Thereaction was purged with N₂ gas for 10 min and then was heated at 90° C.for 0.5 h. The organic layer was carefully separated, absorbed on silicagel and purified by flash column chromatography (0-60% CHCl₃/10% 7 N NH₃in MeOH). The pure fractions were combined, concentrated, the residuesuspended in EtOAc, sonicated and the solids were collected byfiltration. The solid was dried under high vacuo to obtain2-(1H-Imidazol-1-yl)-N-(6-(2-methyloxazol-5-yl)isoquinolin-3-yl)acetamide64 as a pale yellow solid (71.0 mg, 0.213 mmol, 42.6% yield). ¹H NMR(499 MHz, DMSO-d₆) δ ppm 2.53 (3H, s), 5.02 (2H, s), 6.91 (1H, t, J=0.96Hz), 7.20 (1H, t, J=1.23 Hz), 7.67 (1H, s), 7.79 (1H, s), 7.84 (1H, dd,J=8.51, 1.65 Hz), 8.11 (1H, br s), 8.12 (1H, d, J=8.78 Hz), 8.45 (1H,s), 9.15 (1H, s), 10.97 (1H, s); ESIMS found for C₁₈H₁₅N₅O₂ m/z 334.1(M+1).

The following compounds were prepared in accordance with the proceduresdescribed in the above Examples 1-3.

1-Isopropyl-N-(6-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide1

White solid (73.0 mg, 0.165 mmol, 59.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.46 (6H, d, J=6.59 Hz), 1.94-2.13 (6H, m), 2.22 (3H, s),2.87 (2H, br d, J=11.25 Hz), 4.10-4.20 (1H, m), 4.55 (1H, spt, J=6.63Hz), 7.80 (1H, dd, J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.78 Hz), 8.11 (1H,s), 8.12 (1H, s), 8.17 (1H, s), 8.50 (1H, s), 8.54 (1H, s), 8.60 (1H,s), 9.08 (1H, s), 10.48 (1H, s); ESIMS found for C₂₅H₂₉N₇O m/z 444.2(M+1).

N-(6-(1-(Methyl-d3)-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(piperidin-4-yl)-1H-pyrazole-4-carboxamide3

White solid (133.0 mg, 0.329 mmol, 85.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.77 (2H, qd, J=11.98, 3.84 Hz), 2.00 (2H, br dd,J=11.66, 2.06 Hz), 2.24 (1H, br s), 2.60 (2H, td, J=12.21, 2.20 Hz),3.01-3.09 (2H, m), 4.24 (1H, tt, J=11.56, 4.08 Hz), 7.77 (1H, dd,J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.51 Hz), 8.08 (1H, s), 8.10 (1H, d,J=0.82 Hz), 8.16 (1H, s), 8.37 (1H, s), 8.54 (1H, s), 8.60 (1H, s), 9.08(1H, s), 10.49 (1H, s); ESIMS found for C₂₂H₂₀[²H₃]N₇O m/z 405.2 (M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-pyrazole-4-carboxamide4

White solid (184.0 mg, 0.421 mmol, 84.4% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.87-2.00 (2H, m), 2.00-2.12 (4H, m), 2.21 (3H, s), 2.86(2H, br d, J=11.80 Hz), 3.91 (3H, s), 4.17 (1H, tt, J=11.11, 4.25 Hz),7.77 (1H, dd, J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.51 Hz), 8.08 (1H, s),8.10 (1H, s), 8.18 (1H, s), 8.37 (1H, s), 8.54 (1H, s), 8.61 (1H, s),9.08 (1H, s), 10.49 (1H, s); ESIMS found for C₂₃H₂₅N₇O m/z 416.2 (M+1).

N-(6-(1-(Methyl-d₃)-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-pyrazole-4-carboxamide5

White solid (95.0 mg, 0.229 mmol, 74.8% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.89-2.00 (2H, m), 2.00-2.11 (4H, m), 2.21 (3H, s), 2.86(2H, br d, J=11.80 Hz), 4.17 (1H, tt, J=11.11, 4.25 Hz), 7.77 (1H, dd,J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.51 Hz), 8.08 (1H, s), 8.10 (1H, d,J=0.82 Hz), 8.18 (1H, s), 8.37 (1H, d, J=0.82 Hz), 8.54 (1H, s), 8.61(1H, s), 9.08 (1H, s), 10.49 (1H, s); ESIMS found for C₂₃H₂₂[²H₃]N₇O m/z419.2 (M+1).

1-(1-Ethylpiperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide6

White solid (34.0 mg, 0.075 mmol, 32.9% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.04 (3H, br t, J=7.00 Hz), 1.90-2.01 (2H, m), 2.03-2.15(4H, m), 2.40 (2H, br s), 2.98 (2H, br s), 3.91 (3H, s), 4.14-4.26 (1H,m), 7.77 (1H, dd, J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.51 Hz), 8.08 (1H,s), 8.10 (1H, s), 8.18 (1H, s), 8.37 (1H, s), 8.54 (1H, s), 8.61 (1H,s), 9.08 (1H, s), 10.47 (1H, s); ESIMS found for C₂₄H₂₇N₇O m/z 430.0(M+1).

1-(1-(2-Fluoroethyl)piperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide 7

White solid (59.0 mg, 0.132 mmol, 57.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.89-2.01 (2H, m), 2.02-2.09 (2H, m), 2.19-2.28 (2H, m),2.68 (2H, dt, J=28.30, 5.00 Hz), 2.99 (2H, br d, J=11.80 Hz), 3.91 (3H,s), 4.20 (1H, tt, J=11.32, 4.19 Hz), 4.56 (2H, td, J=47.90, 5.00 Hz),7.77 (1H, dd, J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.78 Hz), 8.08 (1H, s),8.10 (1H, s), 8.18 (1H, s), 8.37 (1H, s), 8.54 (1H, s), 8.61 (1H, s),9.08 (1H, s), 10.47 (1H, s); ESIMS found for C₂₄H₂₆FN₇O m/z 448.0 (M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(piperidin-4-yl)-1H-1,2,3-triazole-4-carboxamide11

Beige solid (345.0 mg, 0.857 mmol, 93.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.83-1.95 (2H, m), 2.08 (2H, br dd, J=11.11, 1.78 Hz),2.60-2.68 (2H, m), 3.05-3.12 (2H, m), 3.91 (3H, s), 4.61-4.71 (1H, m),7.80 (1H, dd, J=8.64, 1.51 Hz), 8.06 (1H, d, J=8.78 Hz), 8.11 (1H, s),8.14 (1H, s), 8.37 (1H, s), 8.52 (1H, s), 8.94 (1H, s), 9.10 (1H, s),9.98 (1H, br s); ESIMS found for C₂₁H₂₂N₈O m/z 402.95 (M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-1,2,3-triazole-4-carboxamide12

White solid (140.0 mg, 0.319 mmol, 85.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.05-2.17 (6H, m), 2.23 (3H, s), 2.85-2.90 (2H, m), 3.91(3H, s), 4.54-4.64 (1H, m), 7.81 (1H, dd, J=8.51, 1.65 Hz), 8.06 (1H, d,J=8.51 Hz), 8.11 (1H, s), 8.15 (1H, s), 8.38 (1H, s), 8.52 (1H, s), 8.97(1H, s), 9.10 (1H, s), 9.97 (1H, s); ESIMS found for C₂₂H₂₄N₈O m/z 417.0(M+1).

1-(1-(2-Fluoroethyl)piperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-1,2,3-triazole-4-carboxamide 13

White solid (42.0 mg, 0.094 mmol, 75.4% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.02-2.12 (2H, m), 2.12-2.18 (2H, m), 2.24-2.32 (2H, m),2.69 (2H, dt, J=28.60, 4.70 Hz), 2.98-3.06 (2H, m), 3.91 (3H, s),4.48-4.67 (3H, m), 7.81 (1H, dd, J=8.64, 1.51 Hz), 8.06 (1H, d, J=8.51Hz), 8.12 (1H, s), 8.15 (1H, s), 8.39 (1H, s), 8.52 (1H, s), 8.99 (1H,s), 9.10 (1H, s), 10.01 (1H, s); ESIMS found for C₂₃H₂₅FN₈O m/z 449.2(M+1).

1-(1-Isopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-1H-1,2,3-triazole-4-carboxamide14

White solid (163.0 mg, 0.348 mmol, 82.5% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.01 (6H, d, J=6.59 Hz), 1.97-2.07 (2H, m), 2.12-2.19(2H, m), 2.29-2.36 (2H, m), 2.73-2.82 (1H, m), 2.87-2.96 (2H, m), 3.91(3H, s), 4.52-4.61 (1H, m), 7.81 (1H, dd, J=8.51, 1.37 Hz), 8.06 (1H, d,J=8.51 Hz), 8.11 (1H, s), 8.14 (1H, s), 8.37 (1H, s), 8.52 (1H, s), 8.96(1H, s), 9.10 (1H, s), 9.96 (1H, s); ESIMS found for C₂₄H₂₈N₈O m/z 445.0(M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-2-(4-methylpiperazin-1-yl)oxazole-5-carboxamide15

Yellow solid (37.0 mg, 0.084 mmol, 24.3% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.23 (3H, s), 2.39-2.45 (4H, m), 3.56-3.63 (4H, m), 3.91(3H, s), 7.78 (1H, dd, J=8.51, 1.65 Hz), 7.91 (1H, s), 8.04 (1H, d,J=8.51 Hz), 8.08 (1H, s), 8.11 (1H, s), 8.38 (1H, s), 8.46 (1H, s), 9.08(1H, s), 10.57 (1H, s); ESIMS found for C₂₂H₂₃N₇O₂ m/z 418.2 (M+1).

N-(6-(1-(Methyl-d₃)-1H-pyrazol-4-yl)isoquinolin-3-yl)-2-(piperidin-4-yl)oxazole-4-carboxamide 16

White solid (209.0 mg, 0.516 mmol, 89.9% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.64-1.75 (2H, m), 2.00 (2H, br dd, J=12.90, 2.74 Hz),2.65-2.72 (2H, m), 3.02-3.10 (3H, m), 7.81 (1H, dd, J=8.51, 1.65 Hz),8.06 (1H, d, J=8.51 Hz), 8.12 (1H, s), 8.15 (1H, s), 8.39 (1H, s), 8.50(1H, s), 8.85 (1H, s), 9.09 (1H, s), 9.72 (1H, br s); ESIMS found forC₂₂H₁₉[²H₃]N₆O₂ m/z 406.2 (M+1).

N-(6-(1-(Methyl-d₃)-1H-pyrazol-4-yl)isoquinolin-3-yl)-2-(1-methylpiperidin-4-yl)oxazole-4-carboxamide17

White solid (154.0 mg, 0.367 mmol, 73.3% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.73-1.84 (2H, m), 1.98-2.07 (4H, m), 2.19 (3H, s), 2.78(2H, br d, J=11.25 Hz), 2.88 (1H, tt, J=11.22, 3.74 Hz), 7.81 (1H, dd,J=8.64, 1.51 Hz), 8.06 (1H, d, J=8.51 Hz), 8.12 (1H, s), 8.15 (1H, s),8.38 (1H, s), 8.50 (1H, s), 8.85 (1H, s), 9.09 (1H, s), 9.70 (1H, s);ESIMS found for C₂₃H₂₁[²H₃]N₆O₂ m/z 420.2 (M+1).

2-(3-(Dimethylamino)azetidin-1-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)oxazole-4-carboxamide 18

Off-white solid (17.6 mg, 0.042 mmol, 9.5% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.12 (6H, s), 3.22-3.30 (1H, m), 3.90 (3H, s), 3.96 (2H,dd, J=8.37, 5.35 Hz), 4.16 (2H, t, J=7.68 Hz), 7.80 (1H, dd, J=8.64,1.51 Hz), 8.05 (1H, d, J=8.78 Hz), 8.11 (1H, d, J=0.82 Hz), 8.14 (1H,s), 8.39 (1H, s), 8.39 (1H, s), 8.48 (1H, s), 9.07 (1H, s), 9.43 (1H,s); ESIMS found for C₂₂H₂₃N₇O₂ m/z 418.2 (M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-2-(piperidin-4-yl)oxazole-4-carboxamide19

White solid (818.0 mg, 2.033 mmol, 92.9% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.58-1.69 (2H, m), 1.95 (2H, br dd, J=12.76, 2.61 Hz),2.59 (2H, td, J=11.87, 2.33 Hz), 2.95-3.04 (3H, m), 3.91 (3H, s), 7.81(1H, dd, J=8.51, 1.65 Hz), 8.06 (1H, d, J=8.78 Hz), 8.12 (1H, s), 8.15(1H, s), 8.39 (1H, s), 8.50 (1H, s), 8.84 (1H, s), 9.09 (1H, s), 9.70(1H, br s); ESIMS found for C₂₂H₂₂N₆O₂ m/z 403.0 (M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-2-(1-methylpiperidin-4-yl)oxazole-4-carboxamide20

White solid (83.0 mg, 0.189 mmol, 63.5% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.72-1.85 (2H, m), 1.98-2.07 (4H, m), 2.19 (3H, s), 2.79(2H, br d, J=11.53 Hz), 2.88 (1H, tt, J=11.25, 3.57 Hz), 3.91 (3H, s),7.81 (1H, dd, J=8.51, 1.37 Hz), 8.06 (1H, d, J=8.51 Hz), 8.12 (1H, s),8.15 (1H, s), 8.39 (1H, s), 8.50 (1H, s), 8.85 (1H, s), 9.09 (1H, s),9.71 (1H, s); ESIMS found for C₂₃H₂₄N₆O₂ m/z 416.95 (M+1).

2-(1-(2-Fluoroethyl)piperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)oxazole-4-carboxamide 21

Beige solid (31.0 mg, 0.069 mmol, 27.8% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.73-1.85 (2H, m), 2.01-2.08 (2H, m), 2.16-2.25 (2H, m),2.65 (2H, dt, J=28.60, 5.00 Hz), 2.88-2.97 (3H, m), 3.91 (3H, s), 4.55(2H, dt, J=47.90, 5.00 Hz), 7.81 (1H, dd, J=8.51, 1.65 Hz), 8.06 (1H, d,J=8.51 Hz), 8.12 (1H, s), 8.15 (1H, s), 8.39 (1H, s), 8.50 (1H, s), 8.85(1H, s), 9.09 (1H, s), 9.71 (1H, s); ESIMS found for C₂₄H₂₅FN₆O₂ m/z449.2 (M+1).

2-(1-Isopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)oxazole-4-carboxamide22

White solid (144.0 mg, 0.308 mmol, 82.6% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 0.98 (6H, d, J=6.59 Hz), 1.72 (2H, qd, J=11.76, 3.43 Hz),2.00-2.08 (2H, m), 2.25 (2H, td, J=11.25, 1.92 Hz), 2.66-2.77 (1H, m),2.78-2.85 (2H, m), 2.85-2.92 (1H, m), 3.91 (3H, s), 7.81 (1H, dd,J=8.51, 1.65 Hz), 8.06 (1H, d, J=8.51 Hz), 8.12 (1H, s), 8.15 (1H, s),8.39 (1H, s), 8.50 (1H, s), 8.84 (1H, s), 9.09 (1H, s), 9.70 (1H, s);ESIMS found for C₂₅H₂₈N₆O₂ m/z 445.2 (M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-2-(4-methylpiperazin-1-yl)oxazole-4-carboxamide23

Off-white solid (40.0 mg, 0.096 mmol, 28.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.23 (3H, s), 2.39-2.45 (4H, m), 3.47-3.54 (4H, m), 3.90(3H, s), 7.80 (1H, dd, J=8.51, 1.37 Hz), 8.05 (1H, d, J=8.51 Hz), 8.11(1H, s), 8.14 (1H, s), 8.38 (1H, s), 8.39 (1H, s), 8.49 (1H, s), 9.07(1H, s), 9.45 (1H, s); ESIMS found for C₂₂H₂₃N₇O₂ m/z 418.2 (M+1).

2-(3-(Dimethylamino)azetidin-1-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)thiazole-5-carboxamide 24

White solid (12.1 mg, 0.028 mmol, 6.3% yield). ¹H NMR (499 MHz, DMSO-d₆)δ ppm 2.13 (6H, s), 3.88-3.95 (2H, m), 3.91 (3H, s), 4.11 (2H, t, J=7.96Hz), 7.77 (1H, dd, J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.51 Hz), 8.07 (1H,s), 8.09 (1H, s), 8.33 (1H, s), 8.36 (1H, s), 8.44 (1H, s), 9.08 (1H,s), 10.76 (1H, s); ESIMS found for C₂₂H₂₃N₇OS m/z 434.1 (M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-2-(piperidin-4-yl)thiazole-4-carboxamide 25

White solid (880.0 mg, 2.103 mmol, 72.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.63 (2H, qd, J=11.98, 3.84 Hz), 2.00-2.06 (2H, m), 2.24(1H, br s), 2.62 (2H, td, J=12.08, 1.92 Hz), 2.99-3.08 (2H, m), 3.18(1H, tt, J=11.66, 3.84 Hz), 3.91 (3H, s), 7.81 (1H, dd, J=8.51, 1.37Hz), 8.07 (1H, d, J=8.51 Hz), 8.12 (1H, s), 8.15 (1H, s), 8.39 (1H, s),8.46 (1H, s), 8.54 (1H, s), 9.09 (1H, s), 9.88 (1H, br s); ESIMS foundfor C₂₂H₂₂N₆OS m/z 419.1 (M+1).

N-(6-(1-Methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)-2-(4-methylpiperazin-1-yl)thiazole-4-carboxamide26

Off-white solid (17.8 mg, 0.041 mmol, 4.6% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.23 (3H, s), 2.41-2.47 (4H, m), 3.49-3.55 (4H, m), 3.91(3H, s), 7.76 (1H, dd, J=8.51, 1.65 Hz), 8.03 (1H, d, J=8.51 Hz), 8.07(1H, s), 8.09 (1H, s), 8.35 (1H, s), 8.36 (1H, s), 8.44 (1H, s), 9.08(1H, s), 10.75 (1H, s); ESIMS found for C₂₂H₂₃N₇OS m/z 434.2 (M+1).

2-(1-(2-Fluoroethyl)piperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)thiazole-4-carboxamide 27

Beige solid (25.0 mg, 0.049 mmol, 20.5% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.79 (2H, qd, J=11.98, 3.57 Hz), 2.08-2.15 (2H, m), 2.21(2H, br t, J=11.53 Hz), 2.67 (2H, dt, J=28.30, 5.00 Hz), 2.99 (2H, br d,J=11.53 Hz), 3.10 (1H, tt, J=11.46, 3.77 Hz), 3.91 (3H, s), 4.56 (2H,dt, J=48.10, 5.00 Hz), 7.81 (1H, dd, J=8.51, 1.65 Hz), 8.07 (1H, d,J=8.51 Hz), 8.12 (1H, s), 8.16 (1H, s), 8.39 (1H, s), 8.47 (1H, s), 8.55(1H, s), 9.10 (1H, s), 9.90 (1H, s); ESIMS found for C₂₄H₂₅FN₆OS m/z511.2 (M+1+formic acid).

2-Methyl-N-(6-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)isoquinolin-3-yl)thiazole-5-carboxamide28

Beige solid (33.0 mg, 0.076 mmol, 53.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.97-2.11 (6H, m), 2.22 (3H, s), 2.71 (3H, s), 2.88 (2H,br d, J=11.25 Hz), 4.12-4.20 (1H, m), 7.83 (1H, dd, J=8.51, 1.37 Hz),8.06 (1H, d, J=8.78 Hz), 8.13 (1H, s), 8.13 (1H, br s), 8.49 (1H, s),8.50 (1H, s), 8.68 (1H, s), 9.11 (1H, s), 11.14 (1H, s); ESIMS found forC₂₃H₂₄N₆OS m/z 433.2 (M+1).

N-(6-(1-Methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-1-(piperidin-4-yl)-1H-pyrazole-4-carboxamide29

Beige solid (47.0 mg, 0.117 mmol, 39.1% yield). ¹H NMR (500 MHz,DMSO-d₆) δ ppm 1.71-1.84 (2H, m), 2.00 (2H, br dd, J=11.66, 2.06 Hz),2.59 (2H, td, J=12.28, 2.33 Hz), 3.01-3.08 (2H, m), 4.15 (3H, s), 4.24(1H, tt, J=11.56, 4.08 Hz), 8.03 (1H, dd, J=8.51, 1.37 Hz), 8.14 (1H, d,J=8.51 Hz), 8.17 (1H, s), 8.33 (1H, s), 8.61 (2H, s), 8.75 (1H, s), 9.17(1H, s), 10.57 (1H, s); ESIMS found for C₂₁H₂₂N₈O m/z 403.2 (M+1).

N-(6-(1-Methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-pyrazole-4-carboxamide30

Beige solid (30.0 mg, 0.072 mmol, 69.0% yield). ¹H NMR (500 MHz,DMSO-d₆) δ ppm 1.93-1.99 (2H, m), 2.02-2.11 (4H, m), 2.21 (3H, s), 2.86(2H, br d, J=11.53 Hz), 4.15 (3H, s), 4.15-4.22 (1H, m), 8.03 (1H, dd,J=8.51, 1.37 Hz), 8.14 (1H, d, J=8.51 Hz), 8.19 (1H, s), 8.33 (1H, s),8.61 (1H, s), 8.62 (1H, s), 8.75 (1H, s), 9.17 (1H, s), 10.57 (1H, s);ESIMS found for C₂₂H₂₄N₈O m/z 417.2 (M+1).

1-(1-Ethylpiperidin-4-yl)-N-(6-(1-methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide31

Beige solid (39.0 mg, 0.086 mmol, 26.6% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.02 (3H, t, J=7.14 Hz), 1.89-1.98 (2H, m), 2.01-2.11(4H, m), 2.37 (2H, q, J=7.32 Hz), 2.96 (2H, brd, J=11.53 Hz), 4.15 (3H,s), 4.16-4.23 (1H, m), 8.03 (1H, dd, J=8.51, 1.37 Hz), 8.14 (1H, d,J=8.51 Hz), 8.19 (1H, s), 8.32 (1H, s), 8.61 (1H, s), 8.62 (1H, s), 8.75(1H, s), 9.17 (1H, s), 10.56 (1H, s); ESIMS found for C₂₃H₂₆N₈O m/z431.2 (M+1).

1-(1-Isopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide 32

Beige solid (67.0 mg, 0.0151 mmol, 55.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 0.99 (6H, d, J=6.59 Hz), 1.89 (2H, qd, J=11.89, 3.84 Hz),2.07 (2H, br dd, J=12.08, 1.92 Hz), 2.24-2.32 (2H, m), 2.75 (1H, spt,J=6.54 Hz), 2.88 (2H, br d, J=11.80 Hz), 4.10-4.20 (1H, m), 4.15 (3H,s), 8.03 (1H, dd, J=8.51, 1.65 Hz), 8.14 (1H, d, J=8.51 Hz), 8.18 (1H,s), 8.32 (1H, s), 8.61 (1H, s), 8.61 (1H, s), 8.75 (1H, s), 9.17 (1H,s), 10.55 (1H, s); ESIMS found for C₂₄H₂₈N₈O m/z 445.2 (M+1).

N-(6-(1-Methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-1-(piperidin-4-yl)-1H-1,2,3-triazole-4-carboxamide33

Beige solid (300.0 mg, 0.744 mmol, 69.3% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.88 (2H, qd, J=11.94, 3.98 Hz), 2.08 (2H, br dd,J=11.53, 1.92 Hz), 2.17 (1H, br s), 2.56-2.68 (2H, m), 3.07 (2H, brd,J=12.62 Hz), 4.15 (3H, s), 4.65 (1H, tt, J=11.70, 4.08 Hz), 8.08 (1H,dd, J=8.51, 1.37 Hz), 8.17 (1H, d, J=8.51 Hz), 8.40 (1H, s), 8.59 (1H,s), 8.77 (1H, s), 8.97 (1H, s), 9.19 (1H, s), 10.11 (1H, br s); ESIMSfound for C₂₀H₂₁N₉O m/z 404.2 (M+1).

1-(1-(2-Fluoroethyl)piperidin-4-yl)-N-(6-(1-methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-1H-1,2,3-triazole-4-carboxamide 34

White solid (34.0 mg, 0.076 mmol, 29.8% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.02-2.11 (2H, m), 2.12-2.17 (2H, m), 2.28 (2H, td,J=11.66, 1.92 Hz), 2.69 (2H, dt, J=28.60, 5.00 Hz), 3.02 (2H, br d,J=11.80 Hz), 4.15 (3H, s), 4.57 (2H, dt, J=47.90, 5.00 Hz), 4.58-4.67(1H, m), 8.08 (1H, dd, J=8.51, 1.37 Hz), 8.17 (1H, d, J=8.51 Hz), 8.40(1H, s), 8.59 (1H, s), 8.76 (1H, s), 9.00 (1H, s), 9.19 (1H, s), 10.09(1H, s); ESIMS found for C₂₂H₂₄FN₉O m/z 450.2 (M+1).

1-(1-Isopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-1H-1,2,3-triazole-4-carboxamide 35

Beige solid (88.0 mg, 0.188 mmol, 75.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.01 (6H, d, J=6.59 Hz), 2.01 (2H, qd, J=11.80, 3.57 Hz),2.16 (2H, br d, J=10.70 Hz), 2.34 (2H, br t, J=11.25 Hz), 2.79 (1H, dt,J=12.97, 6.55 Hz), 2.93 (2H, br d, J=11.80 Hz), 4.15 (3H, s), 4.57 (1H,tt, J=11.46, 4.19 Hz), 8.08 (1H, dd, J=8.64, 1.51 Hz), 8.17 (1H, d,J=8.51 Hz), 8.40 (1H, s), 8.59 (1H, s), 8.76 (1H, s), 8.98 (1H, s), 9.19(1H, s), 10.09 (1H, s); ESIMS found for C₂₃H₂₇N₉O m/z 446.25 (M+1).

N-(6-(1-Methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-2-(piperidin-4-yl)oxazole-4-carboxamide 36

Beige solid (280.0 mg, 0.694 mmol, 54.6% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.64 (2H, qd, J=11.89, 3.84 Hz), 1.96 (2H, br dd,J=12.62, 2.74 Hz), 2.60 (2H, td, J=11.87, 2.33 Hz), 2.95-3.05 (3H, m),4.15 (3H, s), 8.08 (1H, dd, J=8.51, 1.65 Hz), 8.17 (1H, d, J=8.78 Hz),8.39 (1H, s), 8.58 (1H, s), 8.77 (1H, s), 8.85 (1H, s), 9.18 (1H, s),9.77 (1H, br s); ESIMS found for C₂₁H₂₁N₇O₂ m/z 404.2 (M+1).

N-(6-(1-Methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)-2-(1-methylpiperidin-4-yl)oxazole-4-carboxamide37

White solid (194.0 mg, 0.465 mmol, 75.0% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.73-1.84 (2H, m), 1.99-2.07 (4H, m), 2.19 (3H, s), 2.79(2H, br d, J=11.53 Hz), 2.88 (1H, tt, J=11.22, 3.74 Hz), 4.15 (3H, s),8.08 (1H, dd, J=8.37, 1.51 Hz), 8.17 (1H, d, J=8.51 Hz), 8.39 (1H, s),8.57 (1H, s), 8.76 (1H, s), 8.86 (1H, s), 9.18 (1H, s), 9.77 (1H, s);ESIMS found for C₂₂H₂₃N₇O₂ m/z 418.2 (M+1).

2-(1-Isopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-1,2,3-triazol-4-yl)isoquinolin-3-yl)oxazole-4-carboxamide 38

Beige solid (55.0 mg, 0.124 mmol, 62.3% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 0.98 (6H, d, J=6.59 Hz), 1.69-1.79 (2H, m), 2.01-2.09(2H, m), 2.26 (2H, td, J=11.25, 2.20 Hz), 2.66-2.76 (1H, m), 2.79-2.85(2H, m), 2.86-2.92 (1H, m), 4.15 (3H, s), 8.08 (1H, dd, J=8.51, 1.65Hz), 8.17 (1H, d, J=8.51 Hz), 8.39 (1H, s), 8.57 (1H, s), 8.76 (1H, s),8.85 (1H, s), 9.18 (1H, s), 9.76 (1H, s); ESIMS found for C₂₄H₂₇N₇O₂ m/z446.2 (M+1).

2-(1-Isopropylpiperidin-4-yl)-N-(6-(1-methyl-1H-pyrazol-4-yl)isoquinolin-3-yl)thiazole-4-carboxamide39

Beige solid (29.0 mg, 0.060 mmol, 26.4% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 0.99 (6H, d, J=6.59 Hz), 1.73 (2H, qd, J=11.98, 3.57 Hz),2.07-2.15 (2H, m), 2.27 (2H, td, J=11.46, 2.06 Hz), 2.73 (1H, quin,J=6.59 Hz), 2.84-2.92 (2H, m), 3.06 (1H, tt, J=11.66, 3.98 Hz), 3.91(3H, s), 7.81 (1H, dd, J=8.51, 1.65 Hz), 8.07 (1H, d, J=8.78 Hz), 8.12(1H, s), 8.15 (1H, s), 8.39 (1H, s), 8.46 (1H, s), 8.54 (1H, s), 9.09(1H, s), 9.89 (1H, s); ESIMS found for C₂₅H₂₈N₆OS m/z 461.2 (M+1).

1-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide40

Off-white solid (24.0 mg, 0.072 mmol, 32.4% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 3.84 (3H, s), 3.90 (3H, s), 7.32 (1H, s), 7.69 (1H, dd,J=8.51, 1.65 Hz), 7.80 (1H, s), 8.04 (1H, s), 8.12 (1H, d, J=8.51 Hz),8.18 (1H, s), 8.47 (1H, s), 8.63 (1H, s), 9.18 (1H, s), 10.59 (1H, s);ESIMS found for C₁₈H₁₆N₆O m/z 333.1 (M+1).

1-Ethyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide41

Off-white solid (55.0 mg, 0.159 mmol, 35.6% yield). ¹H NMR (500 MHz,DMSO-d₆) δ ppm 1.41 (3H, t, J=7.27 Hz), 3.84 (3H, s), 4.19 (2H, q,J=7.41 Hz), 7.32 (1H, d, J=0.82 Hz), 7.69 (1H, dd, J=8.51, 1.65 Hz),7.80 (1H, s), 8.04 (1H, s), 8.12 (1H, d, J=8.51 Hz), 8.19 (1H, s), 8.54(1H, s), 8.63 (1H, s), 9.18 (1H, s), 10.58 (1H, s); ESIMS found forC₁₉H₁₈N₆O m/z 347.15 (M+1).

1-Isopropyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide42

Off-white solid (25.0 mg, 0.069 mmol, 31.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.47 (6H, d, J=6.59 Hz), 3.85 (3H, s), 4.53 (1H, spt,J=6.68 Hz), 7.33 (1H, s), 7.70 (1H, dd, J=8.51, 1.65 Hz), 7.80 (1H, s),7.96 (1H, d, J=1.10 Hz), 8.08 (1H, s), 8.10-8.16 (2H, m), 8.61 (1H, s),9.16 (1H, s), 9.62 (1H, s); ESIMS found for C₂₀H₂₀N₆O m/z 361.2 (M+1).

1-Isopropyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl-1-d)-1H-pyrazole-4-carboxamide43

Dark brown solid (35.0 mg, 0.097 mmol, 67.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.46 (6H, d, J=6.59 Hz), 3.84 (3H, s), 4.55 (1H, spt,J=6.72 Hz), 7.32 (1H, d, J=1.10 Hz), 7.69 (1H, dd, J=8.51, 1.65 Hz),7.80 (1H, s), 8.04 (1H, s), 8.12 (1H, d, J=8.51 Hz), 8.16 (1H, s), 8.60(1H, s), 8.63 (1H, s), 10.55 (1H, s); ESIMS found for C₂₀H₁₉[²H]N₆O m/z362.2 (M+1).

1-Cyclopropyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide44

Off-white solid (72.0 mg, 0.201 mmol, 45.1% yield). ¹H NMR (500 MHz,DMSO-d₆) δ ppm 0.99-1.05 (2H, m), 1.06-1.11 (2H, m), 3.79-3.83 (1H, m),3.84 (3H, s), 7.32 (1H, d, J=1.10 Hz), 7.69 (1H, dd, J=8.51, 1.65 Hz),7.80 (1H, s), 8.03 (1H, d, J=0.82 Hz), 8.12 (1H, d, J=8.78 Hz), 8.13(1H, d, J=0.55 Hz), 8.60 (1H, s), 8.62 (1H, s), 9.18 (1H, s), 10.56 (1H,s); ESIMS found for C₂₀H₁₈N₆O m/z 359.2 (M+1).

1-Isobutyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide45

Off-white solid (73.0 mg, 0.195 mmol, 43.7% yield). ¹H NMR (500 MHz,DMSO-d₆) δ ppm 0.87 (6H, d, J=6.59 Hz), 2.07-2.21 (1H, m), 3.84 (3H, s),3.98 (2H, d, J=7.14 Hz), 7.32 (1H, d, J=0.82 Hz), 7.69 (1H, dd, J=8.51,1.65 Hz), 7.80 (1H, s), 8.04 (1H, d, J=0.82 Hz), 8.12 (1H, d, J=8.51Hz), 8.20 (1H, s), 8.50 (1H, s), 8.63 (1H, s), 9.18 (1H, s), 10.60 (1H,s); ESIMS found for C₂₁H₂₂N₆O m/z 375.2 (M+1).

1-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-pyrazole-3-carboxamide46

Off-white solid (45.0 mg, 0.135 mmol, 60.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 3.85 (3H, s), 3.99 (3H, s), 6.89 (1H, d, J=2.20 Hz), 7.33(1H, s), 7.72 (1H, dd, J=8.51, 1.92 Hz), 7.81 (1H, s), 7.91 (1H, d,J=2.20 Hz), 8.09 (1H, s), 8.13 (1H, d, J=8.51 Hz), 8.60 (1H, s), 9.17(1H, s), 9.64 (1H, s); ESIMS found for C18H₁₆N₆O m/z 333.1 (M+1).

1-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-imidazole-4-carboxamide48

Off-white solid (10.5 mg, 0.032 mmol, 14.2% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 3.76 (3H, s), 3.85 (3H, s), 7.33 (1H, s), 7.70 (1H, dd,J=8.51, 1.65 Hz), 7.80 (1H, s), 7.82 (1H, d, J=1.10 Hz), 7.95 (1H, d,J=1.10 Hz), 8.07 (1H, s), 8.12 (1H, d, J=8.51 Hz), 8.61 (1H, s), 9.16(1H, s), 9.59 (1H, s); ESIMS found for C₁₈H₁₆N₆O m/z 333.1 (M+1).

1-Isopropyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-imidazole-4-carboxamide49

Off-white solid (11.8 mg, 0.033 mmol, 14.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.46 (6H, d, J=6.86 Hz), 3.84 (3H, s), 4.55 (1H, spt,J=6.63 Hz), 7.32 (1H, s), 7.69 (1H, dd, J=8.51, 1.37 Hz), 7.80 (1H, s),8.04 (1H, s), 8.12 (1H, d, J=8.51 Hz), 8.16 (1H, s), 8.60 (1H, s), 8.63(1H, s), 9.18 (1H, s), 10.55 (1H, s); ESIMS found for C₂₀H₂₀N₆O m/z361.2 (M+1).

N-(6-(1-Methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)-1H-imidazole-4-carboxamide50

Off-white solid (18.0 mg, 0.045 mmol, 20.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.94-2.06 (4H, m), 3.39-3.51 (2H, m), 3.85 (3H, s),3.94-4.03 (2H, m), 4.37-4.49 (1H, m), 7.33 (1H, d, J=1.10 Hz), 7.70 (1H,dd, J=8.51, 1.65 Hz), 7.80 (1H, s), 8.00 (1H, d, J=1.10 Hz), 8.08 (1H,s), 8.12 (1H, d, J=8.78 Hz), 8.13 (1H, d, J=1.37 Hz), 8.61 (1H, s), 9.16(1H, s), 9.62 (1H, s); ESIMS found for C₂₂H₂₂N₆O₂ m/z 403.2 (M+1).

1,2-Dimethyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-imidazole-5-carboxamide51

Off-white solid (26.5 mg, 0.077 mmol, 34.3% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.36 (3H, s), 3.83 (3H, s), 3.85 (3H, s), 7.32 (1H, d,J=0.82 Hz), 7.70 (1H, dd, J=8.51, 1.65 Hz), 7.80 (1H, s), 7.95 (1H, s),8.06 (1H, s), 8.12 (1H, d, J=8.51 Hz), 8.61 (1H, s), 9.18 (1H, s), 10.64(1H, s); ESIMS found for C₁₉H₁₈N₆O m/z 347.15 (M+1).

1-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-1,2,4-triazole-3-carboxamide52

Off-white solid (50.9 mg, 0.153 mmol, 68.5% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 3.86 (3H, s), 4.01 (3H, s), 7.34 (1H, d, J=1.10 Hz),7.71-7.77 (1H, m), 7.81 (1H, s), 8.11-8.18 (2H, m), 8.63 (1H, s), 8.73(1H, s), 9.20 (1H, s), 9.87 (1H, s); ESIMS found for C₁₇H₁₅N₇O m/z 334.1(M+1).

2-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)oxazole-4-carboxamide53

Off-white solid (72.0 mg, 0.216 mmol, 48.4% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.53 (3H, s), 3.91 (3H, s), 7.81 (1H, dd, J=8.51, 1.65Hz), 8.05 (1H, d, J=8.51 Hz), 8.11 (1H, s), 8.15 (1H, s), 8.38 (1H, s),8.50 (1H, s), 8.80 (1H, s), 9.09 (1H, s), 9.71 (1H, s); ESIMS found forC₁₈H₁₅N₅O₂ m/z 334.1 (M+1).

2-Isopropyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)oxazole-4-carboxamide54

Beige solid (100.0 mg, 0.277 mmol, 62.1% yield). ¹H NMR (500 MHz,DMSO-d₆) δ ppm 1.35 (6H, d, J=6.86 Hz), 3.13-3.24 (1H, m), 3.85 (3H, s),7.33 (1H, s), 7.74 (1H, br d, J=7.96 Hz), 7.81 (1H, s), 8.11 (1H, s),8.15 (1H, d, J=8.51 Hz), 8.60 (1H, s), 8.84 (1H, s), 9.19 (1H, s), 9.76(1H, s); ESIMS found for C₂₀H₁₉N₅O₂ m/z 362.2 (M+1).

4-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)oxazole-2-carboxamide55

Off-white solid (50.3 mg, 0.151 mmol, 67.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.53 (3H, s), 3.85 (3H, s), 7.33 (1H, d, J=1.10 Hz), 7.73(1H, dd, J=8.51, 1.65 Hz), 7.80 (1H, s), 8.11 (1H, s), 8.14 (1H, d,J=8.51 Hz), 8.60 (1H, s), 8.81 (1H, s), 9.19 (1H, s), 9.78 (1H, s);ESIMS found for C₁₈H₁₅N₅O₂ m/z 334.1 (M+1).

4-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)thiazole-2-carboxamide56

Off-white solid (30.1 mg, 0.086 mmol, 38.6% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.52 (3H, d, J=0.82 Hz), 3.85 (3H, s), 7.34 (1H, d,J=1.10 Hz), 7.76 (1H, dd, J=8.51, 1.65 Hz), 7.78 (1H, d, J=0.82 Hz),7.81 (1H, s), 8.13 (1H, s), 8.16 (1H, d, J=8.51 Hz), 8.58 (1H, s), 9.22(1H, s), 10.04 (1H, br s); ESIMS found for C₁₈H₁₅N₅OS m/z 350.1 (M+1).

2-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)thiazole-4-carboxamide57

Off-white solid (42.4 mg, 0.121 mmol, 54.4% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.79 (3H, s), 3.85 (3H, s), 7.34 (1H, d, J=1.10 Hz), 7.74(1H, dd, J=8.51, 1.65 Hz), 7.81 (1H, s), 8.11 (1H, s), 8.15 (1H, d,J=8.51 Hz), 8.42 (1H, s), 8.64 (1H, s), 9.19 (1H, s), 9.92 (1H, s);ESIMS found for C₁₈H₁₅N₅OS m/z 350.1 (M+1).

5-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1,3,4-oxadiazole-2-carboxamide58

Off-white solid (20.2 mg, 0.060 mmol, 27.1% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.65 (3H, s), 3.86 (3H, s), 7.35 (1H, s), 7.75-7.81 (1H,m), 7.82 (1H, s), 8.13-8.23 (2H, m), 8.55 (1H, s), 9.24 (1H, s), 11.10(1H, s); ESIMS found for C₁₇H₁₄N₆O₂ m/z 335.1 (M+1).

5-Methyl-N-(6-(1-methyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1,3,4-thiadiazole-2-carboxamide59

Off-white solid (10.0 mg, 0.029 mmol, 12.8% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.86 (3H, s), 3.85 (3H, s), 7.34 (1H, s), 7.78 (1H, dd,J=8.51, 1.65 Hz), 7.81 (1H, s), 8.14 (1H, s), 8.17 (1H, d, J=8.78 Hz),8.56 (1H, s), 9.24 (1H, s), 10.72 (1H, s); ESIMS found for C₁₇H₁₄N₆OSm/z 351.1 (M+1).

N-(6-(1,2-Dimethyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1-methyl-1H-pyrazole-4-carboxamide60

Off-white solid (78.3 mg, 0.226 mmol, 35.9% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.40 (3H, s), 3.67 (3H, s), 3.90 (3H, s), 7.13 (1H, s),7.62 (1H, dd, J=8.51, 1.65 Hz), 7.94 (1H, s), 8.11 (1H, d, J=8.78 Hz),8.18 (1H, s), 8.46 (1H, s), 8.61 (1H, s), 9.17 (1H, s), 10.58 (1H, s);ESIMS found for C₁₉H₁₈N₆O m/z 347.1 (M+1).

N-(6-(1,2-Dimethyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1-isopropyl-1H-pyrazole-4-carboxamide61

Off=white solid (29.9 mg, 0.080 mmol, 12.7% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.46 (6H, d, J=6.59 Hz), 2.40 (3H, s), 3.67 (3H, s), 4.55(1H, spt, J=6.68 Hz), 7.13 (1H, s), 7.62 (1H, dd, J=8.37, 1.51 Hz), 7.94(1H, s), 8.11 (1H, d, J=8.51 Hz), 8.16 (1H, s), 8.60 (1H, s), 8.61 (1H,s), 9.17 (1H, s), 10.54 (1H, s); ESIMS found for C₂₁H₂₂N₆O m/z 375.2(M+1).

N-(6-(1,2-Dimethyl-1H-imidazol-5-yl)isoquinolin-3-yl)-2-methylthiazole-5-carboxamide62

Beige solid (72.0 mg, 0.198 mmol, 53.5% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.40 (3H, s), 2.71 (3H, s), 3.67 (3H, s), 7.14 (1H, s),7.66 (1H, dd, J=8.51, 1.65 Hz), 7.97 (1H, s), 8.14 (1H, d, J=8.51 Hz),8.57 (1H, s), 8.69 (1H, s), 9.21 (1H, s), 11.20 (1H, s); ESIMS found forC₁₉H₁₇N₅OS m/z 364.1 (M+1).

N-(6-(2-Methyloxazol-5-yl)isoquinolin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-pyrazole-4-carboxamide63

White solid (16.0 mg, 0.038 mmol, 85.9% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.90-2.00 (2H, m), 2.01-2.12 (4H, m), 2.21 (3H, s), 2.54(3H, s), 2.86 (2H, br d, J=12.08 Hz), 4.13-4.22 (1H, m), 7.81 (1H, s),7.83 (1H, dd, J=8.51, 1.65 Hz), 8.13 (1H, d, J=8.78 Hz), 8.15 (1H, s),8.18 (1H, s), 8.61 (1H, s), 8.62 (1H, s), 9.16 (1H, s), 10.58 (1H, s);ESIMS found for C₂₃H₂₄N₆O₂ m/z 417.2 (M+1).

N-(6-(5-Methyl-1,3,4-thiadiazol-2-yl)isoquinolin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-pyrazole-4-carboxamide65

Off-white solid (27.3 mg, 0.060 mmol, 59.8% yield). ¹H NMR (499 MHz,METHANOL-d₄) δ ppm 2.15 (3H, s), 2.24-2.38 (4H, m), 2.85 (3H, s),2.86-2.93 (2H, m), 3.41 (2H, br d, J=12.62 Hz), 4.50 (1H, tt, J=10.05,5.04 Hz), 8.05-8.09 (1H, m), 8.09-8.12 (1H, m), 8.14 (1H, s), 8.34 (1H,s), 8.42 (1H, s), 8.60 (1H, s), 9.10 (1H, s); ESIMS found for C₂₂H₂₃N₇OSm/z 434.1 (M+1).

1-(1-Isopropylpiperidin-4-yl)-N-(6-(5-methyl-1,3,4-thiadiazol-2-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide 66

White solid (173.0 mg, 0.374 mmol, 54.4% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 0.99 (6H, d, J=6.59 Hz), 1.89 (2H, qd, J=11.71, 3.57 Hz),2.07 (2H, br d, J=12.62 Hz), 2.28 (2H, br t, J=10.84 Hz), 2.75 (1H, dt,J=13.10, 6.48 Hz), 2.83 (3H, s), 2.88 (2H, br d, J=11.53 Hz), 4.11-4.20(1H, m), 8.11 (1H, dd, J=8.64, 1.23 Hz), 8.18 (1H, s), 8.22 (1H, d,J=8.51 Hz), 8.48 (1H, s), 8.62 (1H, s), 8.72 (1H, s), 9.27 (1H, s),10.64 (1H, s); ESIMS found for C₂₄H₂₇N₇OS m/z 462.2 (M+1).

N-(6-(5-Methyl-1,3,4-thiadiazol-2-yl)isoquinolin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-1,2,3-triazole-4-carboxamide67

Off-white solid (15.0 mg, 0.033 mmol, 39.4% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 2.02-2.16 (6H, m), 2.23 (3H, s), 2.84 (3H, s), 2.88 (2H,br d, J=7.68 Hz), 4.55-4.63 (1H, m), 8.16 (1H, dd, J=8.51, 1.65 Hz),8.25 (1H, d, J=8.51 Hz), 8.57 (1H, s), 8.71 (1H, s), 9.00 (1H, s), 9.29(1H, s), 10.19 (1H, s); ESIMS found for C₂₁H₂₂NOS m/z 435.2 (M+1).

N-(6-(5-Methyl-1,3,4-thiadiazol-2-yl)isoquinolin-3-yl)-2-(piperidin-4-yl)oxazole-4-carboxamide 68

Off-white solid (171.0 mg, 0.407 mmol, 70.6% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.61-1.73 (2H, m), 1.95-2.03 (2H, m), 2.60-2.69 (2H, m),2.84 (3H, s), 2.99-3.08 (3H, m), 8.16 (1H, dd, J=8.51, 1.65 Hz), 8.25(1H, d, J=8.78 Hz), 8.56 (1H, s), 8.69 (1H, s), 8.87 (1H, s), 9.28 (1H,s), 9.87 (1H, br s); ESIMS found for C₂₁H₂₀N₆O₂S m/z 421.2 (M+1).

N-(6-(5-Methyl-1,3,4-thiadiazol-2-yl)isoquinolin-3-yl)-2-(1-methylpiperidin-4-yl)oxazole-4-carboxamide69

Off-white solid (114.0 mg, 0.262 mmol, 74.0% yield). ¹H NMR (499 MHz,DMSO-d₆) δ ppm 1.73-1.84 (2H, m), 1.98-2.08 (4H, m), 2.19 (3H, s), 2.79(2H, br d, J=11.25 Hz), 2.84 (3H, s), 2.88 (1H, tt, J=11.18, 3.77 Hz),8.16 (1H, dd, J=8.51, 1.65 Hz), 8.25 (1H, d, J=8.78 Hz), 8.56 (1H, s),8.69 (1H, s), 8.87 (1H, s), 9.28 (1H, s), 9.86 (1H, s); ESIMS found forC₂₂H₂₂N₆O₂S m/z 435.2 (M+1).

1-Isopropyl-N-(6-(1-isopropyl-1H-imidazol-5-yl)isoquinolin-3-yl)-1H-pyrazole-4-carboxamide70

White solid (5.6 mg, 0.014 mmol, 5.2% yield). ¹H NMR (500 MHz, DMSO-d₆)δ ppm 1.44 (6H, d, J=6.59 Hz), 1.46 (6H, d, J=6.86 Hz), 4.56 (2H, dq,J=12.86, 6.60 Hz), 7.14 (1H, d, J=1.10 Hz), 7.57 (1H, dd, J=8.37, 1.51Hz), 7.93 (1H, s), 8.03 (1H, s), 8.15 (1H, d, J=8.51 Hz), 8.17 (1H, s),8.61 (1H, s), 8.62 (1H, s), 9.21 (1H, s), 10.58 (1H, s); ESIMS found forC₂₂H₂₄N₆O m/z 389.2 (M+1).

Example 4

The screening assay for Wnt activity is described as follows. Reportercell lines can be generated by stably transducing cancer cell lines(e.g., colon cancer) or primary cells (e.g., IEC-6 intestinal cells)with a lentiviral construct that includes a Wnt-responsive promoterdriving expression of the firefly luciferase gene.

SW480 colon carcinoma cells were transduced with a lentiviral vectorexpressing luciferase with a human Sp5 promoter consisting of a sequenceof eight TCF/LEF binding sites. SW480 cells stably expressing theSp5-Luc reporter gene and a hygromycin resistance gene were selected bytreatment with 150 μg/mL of hygromycin for 7 days. These stablytransduced SW480 cells were expanded in cell culture and used for allfurther screening activities. Each compound was dissolved in DMSO as a10 mM stock and used to prepare compound source plates. Serial dilution(1:3, 10-point dose-response curves starting from 10 μM) and compoundtransfer was performed using the ECHO 550 (Labcyte, Sunnyvale, Calif.)into 384-well white solid bottom assay plates (Greiner Bio-One) withappropriate DMSO backfill for a final DMSO concentration of 0.1%. ForSp5-Luc reporter gene assays, the cells were plated at 4,000 cells/wellin 384-well plates with a DMEM medium containing 1% fetal bovine serum,and 1% Penicillin-Streptomycin and incubated for 36 to 48 hours at 37°C. and 5% CO₂. Following incubation, 15 μl of BriteLite Plusluminescence reagent (Perkin Elmer) was added to each well of the384-well assay plates. The plates were placed on an orbital shaker for 2min and then luminescence was quantified using the Envision (PerkinElmer) plate reader. Readings were normalized to DMSO only treatedcells, and normalized activities were utilized for EC₅₀ calculationsusing the dose-response log (inhibitor) vs. response −variable slope(four parameters) nonlinear regression feature available in GraphPadPrism 5.0 (or Dotmatics). For EC₅₀ of >10 μM, the percent inhibition at10 M is provided.

Table 2 shows the measured activity for representative compounds ofFormula I as described herein.

TABLE 2 Compound EC₅₀ (μM) 1 0.387 2 0.027 3 0.023 4 0.014 5 0.022 60.015 7 0.035 8 0.011 9 0.039 10 0.055 11 0.104 12 0.105 13 0.120 140.095 15 0.103 16 0.115 17 0.111 18 1.043 19 0.106 20 0.107 21 0.108 220.180 23 1.017 24 1.319 25 0.546 26 1.754 27 0.300 28 1.165 29 0.030 300.014 31 0.021 32 0.033 33 0.071 34 0.230 35 0.111 36 0.114 37 0.105 380.555 39 0.070 40 0.109 41 0.037 42 0.602 43 0.048 44 0.037 45 0.055 461.246 47 0.556 48 1.156 49 0.035 50 0.100 51 0.466 52 2.184 53 1.891 542.126 55 1.119 56 2.310 57 2.546 58 1.311 59 1.890 60 0.121 61 0.063 620.110 63 0.034 64 1.153 65 0.030 66 0.034 67 0.463 68 0.122 69 2.153 700.115

Example 5

Representative compounds were screened using the assay procedure forDYRK1A kinase activity as described below.

Each compound was dissolved in DMSO as a 10 mM stock and used to preparecompound source plates. Serial dilution (1:3, 11-point dose-responsecurves from 10 M to 0.00016 M) and compound transfer was performed usingthe ECHO 550 (Labcyte, Sunnyvale, Calif.) into 1536-well black-walledround bottom plates (Corning).

The DYRK1A kinase assay was run using the Ser/Thr 18 peptide Z-lyteassay kit according to manufacturer's instructions (Life Technologies—aDivision of Thermo-Fisher). This is a non-radioactive assay usingfluorescence resonance energy transfer (FRET) between coumarin andfluorescein to detect kinase activity which is represented as a ratio ofcoumarin emission/fluorescein emission.

Briefly, recombinant DYRK1A kinase, ATP and Ser/Thr peptide 18 wereprepared in 1× Kinase buffer to final concentrations of 0.19 μg/mL, 30μM, and 4 μM respectively. The mixture was allowed to incubate with therepresentative compounds for one hour at room temperature. All reactionswere performed in duplicate. Unphosphorylated (“0% Control”) andphosphorylated (“100% control”) forms of Ser/Thr 18 served as controlreactions. Additionally, an 11-point dose-response curve ofStaurosporine (1 uM top) was run to serve as a positive compoundcontrol.

After incubation, Development Reagent A was diluted in DevelopmentBuffer then added to the reaction and allowed to further incubate forone hour at room temperature. The plate was read at Ex 400 Em 455 todetect the coumarin signal and Ex 400 Em 520 to measure the signal(EnVision Multilabel Plate Reader, PerkinElmer).

The Emission ratio (Em) was calculated as a ratio of the coumarin (C)emission signal (at 445 nm)/Fluorescein (F) emission signal (at 520 nm).The percent phosphorylation was then calculated using the followingformula: [1−((Em ratio×F100%)−C100%)/((C0%−C100%)+(Emratio×(F100%−F0%)))]. Dose-response curves were generated and inhibitoryconcentration (IC₅₀) values were calculated using non-linear regressioncurve fit in the Dotmatics' Studies Software (Bishops Stortford, UK).

Table 3 shows the measured activity for representative compounds ofFormula I as described herein.

TABLE 3 Compound EC₅₀ (μM) 1 0.039 2 0.002 3 0.003 4 0.002 5 0.002 60.003 7 0.003 8 0.001 9 0.002 10 0.003 11 0.007 12 0.009 13 0.005 140.006 15 0.008 16 0.016 17 0.016 18 0.015 19 0.012 20 0.015 21 0.016 220.016 23 0.018 24 0.002 25 0.039 26 0.003 27 0.014 28 0.024 29 0.003 300.003 31 0.003 32 0.003 33 0.007 34 0.006 35 0.010 36 0.019 37 0.024 380.180 39 0.026 40 0.004 41 0.002 42 0.023 43 0.002 44 0.001 45 0.001 460.005 47 0.003 48 0.018 49 0.002 50 0.021 51 0.004 52 0.009 53 0.014 540.018 55 0.007 56 0.005 57 0.006 58 0.009 59 0.004 60 0.004 61 0.005 620.002 63 0.002 64 0.004 65 0.006 66 0.004 67 0.031 68 0.074 69 0.527 700.009

Example 6

Representative compounds were screened using the assay procedure forGSK3β kinase activity as described below.

Each compound is dissolved in DMSO as a 10 mM stock and used to preparecompound source plates. Serial dilution (1:3, 11-point dose-responsecurves from 10 M to 0.0003 μM) and compound transfer was performed usingthe ECHO 550 (Labcyte, Sunnyvale, Calif.) into 1536-well black-walledround bottom plates (Corning).

The GSK3β kinase assay is run using the Ser/Thr 09 peptide Z-lyte assaykit according to manufacturer's instructions (Life Technologies—aDivision of Thermo-Fisher). This is a non-radioactive assay usingfluorescence resonance energy transfer (FRET) between coumarin andfluorescein to detect kinase activity which is represented as ratio ofcoumarin emission/fluorescein emission.

Briefly, recombinant GSK3β kinase, ATP and Ser/Thr peptide 09 areprepared in 1× Kinase buffer to final concentrations of 0.04 μg/mL, 46μM, and 4 μM respectively. The mixture is allowed to incubate with therepresentative compounds for one hour at room temperature. All reactionswere performed in duplicate. Unphosphorylated (“0% Control”) andphosphorylated (“100% control”) forms of Ser/Thr 18 serve as controlreactions.

After incubation, diluted Development Buffer is added to the reactionand allowed to further incubate for one hour at room temperature. Theplate is read at Ex 400 Em 455 to detect the coumarin signal and Ex 400Em 520 to measure the signal (EnVision Multilabel Plate Reader,PerkinElmer).

The Emission ratio (Em) is calculated as a ratio of the coumarin (C)emission signal (at 445 nm)/Fluorescein (F) emission signal (at 520 nm).The percent phosphorylation is then calculated using the followingformula: [1−((Em ratio×F100%)−C100%)/((C0%−C100%)+(Emratio×(F100%−F0%)))].

Dose-response curves are generated and inhibitory concentration (IC₅₀)values are calculated using non-linear regression curve fit in theDotmatics' Studies Software (Bishops Stortford, UK).

Table 4 shows the activity of representative compounds of Formula I asprovided herein.

TABLE 4 Compound EC₅₀ (μM) 1 9.990 2 1.040 3 >10 4 1.480 5 2.063 6 1.7047 1.495 8 0.780 9 1.656 10 4.175 11 9.990 12 3.634 13 2.082 14 2.747 159.515 16 0.769 17 2.934 18 1.263 19 0.979 20 3.242 21 1.326 22 2.586 232.179 24 1.755 25 3.058 26 2.189 27 3.605 28 >10 29 0.230 30 0.322 310.286 32 0.349 33 0.316 34 0.395 35 0.295 36 0.458 37 0.342 38 1.350 393.708 40 1.826 41 0.850 42 9.990 43 3.071 44 1.655 45 1.194 46 1.769 470.029 48 3.376 49 1.335 50 4.491 51 0.641 52 5.034 53 0.821 54 1.921 550.986 56 9.990 57 1.614 58 4.336 59 1.415 60 4.014 61 6.789 62 0.450 630.611 64 0.009 65 0.628 66 0.390 67 2.339 68 0.666 69 6.769 70 7.080

Example 7

Representative compounds were screened using the assay procedure for tauphosphorylation activity described below.

SH-SY5Y cells (human neuroblastoma) were cultured in DMEM/F-12 mediumsupplemented with 15% FBS, Non-essential Amino Acid andPenicillin/Streptomycin. Two days before treatment, cells were seededonto 96 well plates at 5×10⁴ cells/well.

The above synthesized compounds were screened using the cell assayprocedure to assess decrease Tau phosphorylation at Ser396 (pSer396)described below.

DMSO-resuspended compounds were dispensed to 8 wells as a serialtitration from 10 μM to 4.6 nM final in medium and cells were exposedovernight (16-18 h) in a humidified incubator at 36.6c before harvest.Wells were visually checked for cell death or change in morphology andsupernatants were tested for cytotoxicity by measurement of lactatedehydrogenase release (LDH, CytoToxOne kit, Promega) if necessary. Ascontrols, commercially available DYRK1A inhibitors, Harmine and Indywhich were shown to have good DYRK1A inhibition in the kinase assay withno CDK1 activity (EC₅₀ 18 and 53 nM respectively, 6 μM for CDK1) butweak EC₅₀ in the Tau assay >10 μM.

Cells were lysed with RIPA buffer complemented with phosphatase andprotease inhibitors then lysates were spun down at 12,000 g for 10 minto remove any cellular debris. Lysates are then either directly testedfor pSer396 by ELISA (Life Technology, Kit KHB7031) or loaded on NuPageBis-Tris gels for western blot analysis. Colorimetric detection of ELISAsignal is performed by Cytation3 plate reader (Biotek) and thechemiluminescence signal for HRP-linked antibodies used in westernblotting is detected using a Carestream Image Station. The same pSer396antibody is used for detection of pTau in both assays.

Blot densitometry for pSer396 and β-actin were analyzed using ImageJ(NIH) and pSer396 Tau ELISA signal was used to plot, draw the curvefitting, and determine each compounds EC₅₀ in Prism (GraphPad).

Table 5 shows the activity of representative compounds as providedherein.

TABLE 5 Compound EC₅₀ (μM) 2 0.099 8 0.027 10 0.064 27 >10 28 2.100 620.193 64 0.085

Example 8

Representative compounds were screened using the assay procedure toassess the effect on cell viability as described below.

SW480 colon carcinoma cells were transduced with a lentiviral vectorexpressing luciferase with a human Sp5 promoter consisting of a sequenceof eight TCF/LEF binding sites. SW480 cells stably expressing theSp5-Luc reporter gene and a hygromycin resistance gene were selected bytreatment with 150 pg/mL of hygromycin for 7 days. These stablytransduced SW480 cells were expanded in cell culture and used for allfurther screening activities. Each compound was dissolved in DMSO as a10 mM stock and used to prepare compound source plates. Serial dilution(1:3, 8-point dose-response curves from 10 μM to 0.0045 μM) and compoundtransfer was performed using the ECHO 550 (Labcyte, Sunnyvale, Calif.)into 384-well white solid bottom assay plates (Greiner Bio-One) withappropriate DMSO backfill for a final DMSO concentration of 0.1%.

For the Cell Viability Assays, the cells were plated at 2,000 cells/wellin 384-well plates with a DMEM medium containing 1% fetal bovine serum,and 1% Penicillin-Streptomycin and incubated for four days hours at 37°C. and 5% CO₂. Eight replicates of DMSO-treated cells served as controlsand cells treated with compound were performed in duplicate.

After incubation, 10 μL of CellTiter-Glo (Promega) was added to eachwell allowed to incubate for approximately 12 minutes. This reagent“results in cell lysis and generation of a luminescent signalproportional to the amount of ATP present. The amount of ATP is directlyproportional to the number of cells present in culture, in agreementwith previous reports. The CellTiter-Glo® Assay generates a “glow-type”luminescent signal, produced by the luciferase reaction (Promega.com)”.

After incubation, the plates were read at Ex 560 nm Em 590 nm (Cytation3, BioTek). Dose-response curves were generated and EC₅₀ concentrationvalues were calculated using non-linear regression curve fit in theGraphPad Prism (San Diego, Calif.) or Dotmatics' Studies Software(Bishops Stortford, UK). For EC₅₀ of >10 μM, the percent inhibition at10 μM is provided.

Table 6 shows the activity of representative compounds of Formula I asprovided herein.

TABLE 6 Compound EC₅₀ (μM) 1 0.608 2 0.038 3 0.038 4 0.035 5 0.034 60.032 7 0.077 8 0.059 9 0.104 10 0.105 11 0.273 12 0.470 13 0.485 140.240 15 0.146 16 0.113 17 0.128 18 3.909 19 0.300 20 0.427 21 1.259 220.782 23 3.730 24 1.642 25 0.876 26 3.412 27 3.651 28 0.340 29 0.060 300.032 31 0.039 32 0.044 33 0.257 34 0.664 35 0.306 36 0.399 37 0.179 381.627 39 1.192 40 0.442 41 0.102 42 0.999 43 0.083 44 0.094 45 0.111 462.178 47 0.607 48 3.553 49 0.073 50 0.833 51 0.844 52 3.609 53 3.197 543.731 55 3.067 56 3.073 57 1.807 58 2.956 59 3.320 60 0.405 61 0.111 620.023 63 0.058 64 0.836 65 0.048 66 0.046 67 0.899 68 0.414 69 1.291 700.132

Example 9

Representative compounds were screened using primary human fibroblasts(derived from IPF patients) treated with TGF-β1 to determine theirability to inhibit the fibrotic process.

Human Fibroblast Cell Culture:

Primary human fibroblasts derived from IPF patients (LL29 cells)[¹Xiaoqiu Liu, et. al., “Fibrotic Lung Fibroblasts Show BluntedInhibition by cAMP Due to Deficient cAMP Response Element-BindingProtein Phosphorylation”, Journal of Pharmacology and ExperimentalTherapeutics (2005), 315(2), 678-687; ²Watts, K. L., et. al., “RhoAsignaling modulates cyclin D1 expression in human lung fibroblasts;implications for idiopathic pulmonary fibrosis”, Respiratory Research(2006), 7(1), 88] were obtained from American Type Culture Collection(ATCC) and expanded in F12 medium supplemented with 15% Fetal BovineSerum and 1% Penicillin/Streptomycin.

Compound Screening:

Each compound was dissolved in DMSO as a 10 mM stock and used to preparecompound source plates. Serial dilution (1:2, 11-point dose-responsecurves from 10 μM to 0.94 nM) and compound transfer was performed usingthe ECHO 550 (Labcyte, Sunnyvale, Calif.) into 384-well clear bottomassay plates (Greiner Bio-One) with appropriate DMSO backfill for afinal DMSO concentration of 0.1%. LL29 cells were plated at 1,500cells/well in 70 μL/well F12 medium supplemented with 1% Fetal BovineSerum. TGF-β1 (Peprotech; 20 ng/mL) was added to the plates to inducefibrosis (ref. 1 and 2 above). Wells treated with TGF-β1 and containingDMSO were used as positive control, and cells with only DMSO werenegative control. Cells were incubated at 37° C. and 5% CO₂ for 4 days.Following incubation for 4 days, SYTOX green nucleic acid stain (LifeTechnologies [Thermo Fisher Scientific]) was added to the wells at afinal concentration of 1 μM and incubated at room temperature for 30min. Cells were then fixed using 4% formaldehyde (Electron MicroscopySciences), washed 3 times with PBS followed by blocking andpermeabilization using 3% Bovine Serum Albumin (BSA; Sigma) and 0.3%Triton X-100 (Sigma) in PBS. Cells were then stained with antibodyspecific to α-smooth muscle actin (αSMA; Abcam) (ref. 1 and 2 above) in3% Bovine Serum Albumin (BSA; Sigma) and 0.3% Triton X-100 (Sigma) inPBS, and incubated overnight at 4° C. Cells were then washed 3 timeswith PBS, followed by incubation with Alexa Flor-647 conjugatedsecondary antibody (Life Technologies [Thermo Fisher Scientific]) andDAPI in 3% Bovine Serum Albumin (BSA; Sigma) and 0.3% Triton X-100(Sigma) in PBS at room temperature for 1 hour. Cells were then washed 3times with PBS and plates were sealed for imaging. αSMA staining wasimaged by excitation at 630 nm and emission at 665 nm and quantifiedusing the Compartmental Analysis program on the CellInsight CX5 (ThermoScientific). Dead or apoptotic cells were excluded from analysis basedon positive SYTOX green staining. % of total cells positive for αSMAwere counted in each well and normalized to the average of 11 wellstreated with TGF-β1 on the same plate using Dotmatics' Studies Software.The normalized averages (fold change over untreated) of 3 replicatewells for each compound concentration were used to create dose-responsescurves and EC₅₀ values were calculated using non-linear regression curvefit in the Dotmatics' Studies Software. For EC₅₀ of >10 μM, the percentinhibition at 10 μM is provided.

Table 7 shows the activity of representative compounds of Formula I asprovided herein.

TABLE 7 Compound EC₅₀ (μM) 1 0.953 2 0.093 3 0.074 4 0.094 5 0.084 60.069 7 0.047 8 0.072 9 >10 (33.4%) 10 0.062 11 0.607 12 0.983 13 0.26014 0.410 15 0.279 16 0.312 17 0.173 18 0.481 19 0.170 20 2.000 21 9.61822 2.911 23 0.325 24 2.749 25 1.190 26 4.698 27 2.525 28 0.381 29 0.13530 0.083 31 0.298 32 0.203 33 0.562 34 0.642 35 0.635 36 0.596 37 0.24838 >10 (3.9%)  39 3.039 40 >10 (23.2%) 41 0.122 42 2.502 43 0.151 440.115 45 0.088 46 1.528 47 0.094 48 >10 (6.7%)  49 0.086 50 1.183 51 >10(6.5%)  52 >10 (33.8%) 53 2.421 54 3.209 55 3.811 56 3.192 57 2.827 581.281 59 1.423 60 0.770 61 0.318 62 0.644 63 0.113 64 0.187 65 0.219 660.169 67 4.362 68 2.583 69 >10 (43.9%) 70 0.249

Example 10

Representative compounds were screened using the following assayprocedure to determine their ability to inhibit IL-6 and thereforedemonstrate their anti-inflammatory properties.

Human Peripheral Blood Mononuclear Cells:

Fresh Normal PB MNC (Catalog # PB001, AllCells, Alameda, Calif.) wereshipped overnight at 4° C. and resuspended in Roswell Park MemorialInstitute (RPMI) 1640 Medium, with GlutaMAX Supplement (Catalog#61870127, ThermoFisher Scientific, Waltham, Mass.) supplemented with 1%Penicillin-Streptomycin (Catalog #15140163, ThermoFisher Scientific,Waltham, Mass.) and 1% fetal bovine serum (FBS) (Catalog #16140089,ThermoFisher Scientific, Waltham, Mass.) assay media.

Compound Screening:

Fresh normal human peripheral blood mononuclear cells (huPBMCs) wereresuspended in 1% FBS-RPMI assay media with 1% Penicillin-Streptomycin1% to a cell concentration of 1×10e6 cells/mL. Each compound wasdissolved in DMSO (Catalog # D8418-100 ml, Sigma-Aldrich, St. Louis,Mo.) as a 10 mM stock and used to prepare compound source plates. Serialdilution (1:3, 10-point dose-response curves starting from 10 M) andcompound transfer was performed using the ECHO 550 (Labcyte, Sunnyvale,Calif.) into 384-well white Proxiplate-Plus assay plates (Catalog#6008289, PerkinElmer, Shelton, Conn.) with appropriate DMSO backfillfor a final DMSO concentration of 0.25%. huPBMCs were plated at 5000cells/well in the 384-well Proxiplate-Plus assay plates and incubated at37° C.-5% CO₂ for 2 hours. 50 ng/mL of Lipopolysaccharides fromEscherichia coli 0111:B4 (Catalog #L5293-2ML, Sigma-Aldrich, St. Louis,Mo.) was added after 2 hours and cells were incubated for another 22hours at 37° C.-5% CO₂. After 22 hour incubation, a mixture of anti-IL6XL665 and anti-IL-6 Cryptate diluted in reconstitution buffer (Catalog#62IL6PEC, Cisbio Inc., Bedford, Mass.) was added to each well.Following incubation for 3 hours at room temperature, HomogeneousTime-Resolved Fluorescence (HTRF) was measured using the Envision(Perkin Elmer, Shelton, Conn.) at 665 nm and 620 nM. The ratio offluorescence at 665 nm to 620 nm was used as a readout for IL-6quantification. All samples were processed in duplicate. Readings werenormalized to DMSO treated cells and normalized activities were utilizedfor EC₅₀ calculations. EC₅₀ was determined using software generated byDotmatics Limited (Windhill Bishops Stortford Herts, UK) using theLevenberg-Marquardt 4 parameter fitting procedure with finite differentgradients. For EC₅₀ of >10 μM, the percent inhibition at 10 μM isprovided.

Table 8 shows the activity of representative compounds of Formula I asprovided herein.

TABLE 8 Compound EC₅₀ (μM) 1 3.444 2 0.114 3 1.042 4 0.089 5 0.218 60.129 7 0.145 8 0.504 9 1.099 10 0.277 11 2.627 12 3.750 13 3.386 14 >10(46.0%) 15 0.754 16 1.070 17 6.203 18 >10 (9.0%)  19 0.695 20 3.68721 >10 (27.2%) 22 >10 (27.8%) 23 >10 (5.5%)  24 3.020 25 2.581 26 >10(40.3%) 27 >10 (34.5%) 28 9.975 29 0.366 30 0.261 31 0.688 32 0.437 330.852 34 1.102 35 2.111 36 2.545 37 2.655 38 >10 (5.3%)  39 7.589 400.751 41 0.618 42 >10 (37.1%) 43 0.251 44 0.356 45 0.414 46 >10 (46.1%)47 0.388 48 >10 (17.8%) 49 0.285 50 3.280 51 >10 (10.9%) 52 >10 (1.1%) 53 >10 (47.5%) 54 6.513 55 9.475 56 8.919 57 >10 (19.6%) 58 >10 (9.5%) 59 >10 (0%)   60 1.580 61 0.566 62 2.759 63 0.298 64 0.427 65 0.361 660.893 67 >10 (8.3%)  68 2.635 69 >10 (2.3%)  70 1.598

What is claimed is:
 1. A compound, or a pharmaceutically acceptable saltthereof, of Formula I:

wherein: R¹, R², R⁴, and R⁵ are H; R³ is a 5-membered heteroaryloptionally substituted with 1-4 R³⁶; R⁶ is selected from the groupconsisting of

and

R⁷ is selected from the group consisting of -heterocyclyl optionallysubstituted with 1-10 R⁴⁴ and -carbocyclyl optionally substituted with1-12 R⁴⁵; R¹⁵ is selected from the group consisting of -heterocyclyloptionally substituted with 1-10 R⁴⁴ and -carbocyclyl optionallysubstituted with 1-12 R⁴⁵; each R³⁶ is independently selected from thegroup consisting of H, unsubstituted —(C₁₋₅ alkyl), unsubstituted —(C₂₋₅alkenyl), unsubstituted —(C₂₋₅ alkynyl), and unsubstituted —(C₁₋₅haloalkyl); each R⁴³ is independently halide; each R⁴⁴ is independentlyselected from the group consisting of unsubstituted —(C₁₋₅ alkyl),unsubstituted —(C₂₋₅ alkenyl), unsubstituted —(C₂₋₅ alkynyl),unsubstituted —(C₁₋₅ haloalkyl), and -carbocyclyl optionally substitutedwith 1-12 R⁴³; and each R⁴⁵ is independently halide.
 2. The compound ofclaim 1, wherein R³ is selected from the group consisting of pyrazolyl,imidazolyl, triazolyl, thiadiazolyl, and oxazolyl, each optionallysubstituted with 1-4 R³⁶.
 3. The compound of claim 2, wherein R³ isselected from the group consisting of pyrazol-4-yl, imidazol-5-yl,1,2,3-triazol-4-yl, thiadiazol-2-yl, and oxazol-5-yl, each optionallysubstituted with 1-4 R³⁶.
 4. The compound of claim 3, wherein R³ is apyrazol-4-yl substituted with one —(C₁₋₃ alkyl).
 5. The compound ofclaim 3, wherein R³ is a imidazol-5-yl substituted with one —(C₁₋₃alkyl).
 6. The compound of claim 3, wherein R³ is a imidazol-5-ylsubstituted with two —(C₁₋₃ alkyl).
 7. The compound of claim 3, whereinR³ is a 1,2,3-triazol-4-yl substituted with one —(C₁₋₃ alkyl).
 8. Thecompound of claim 3, wherein R³ is an unsubstituted thiadiazol-2-yl. 9.The compound of claim 3, wherein R³ is a thiadiazol-2-yl substitutedwith one —(C₁₋₃ alkyl).
 10. The compound of claim 3, wherein R³ is anunsubstituted oxazol-5-yl.
 11. The compound of claim 3, wherein R³ is aoxazol-5-yl substituted with one —(C₁₋₃ alkyl).
 12. The compound ofclaim 3, wherein R is


13. The compound of claim 3, wherein R⁶ is


14. The compound of claim 12, wherein R⁷ is selected from the groupconsisting of piperidinyl, piperazinyl, morpholinyl, andtetrahydropyranyl, wherein each is optionally substituted with oneunsubstituted —(C₁₋₃ alkyl).
 15. The compound of claim 13, wherein R¹⁵is selected from the group consisting of piperidinyl, piperazinyl,morpholinyl, and tetrahydropyranyl, wherein each is optionallysubstituted with one unsubstituted —(C₁₋₃ alkyl).
 16. The compound ofclaim 1, wherein the compound of Formula I is selected from the groupconsisting of:

or a pharmaceutically acceptable salt thereof.
 17. A pharmaceuticalcomposition comprising a therapeutically effective amount of a acompound of claim 1, or a pharmaceutically acceptable salt thereof, anda pharmaceutically acceptable excipient.
 18. A method of treating adisorder or disease in a patient, wherein the disorder or disease isselected from the group consisting of: chronic inflammation, systemicinflammation, diabetes, cancer, pulmonary fibrosis, idiopathic pulmonaryfibrosis (IPF), degenerative disc disease, bone/osteoporotic fractures,a bone or cartilage disease, a neurological condition/disorder/disease,osteoarthritis, lung disease, a fibrotic disorder, the method comprisingadministering to the patient a therapeutically effective amount a of acompound of claim 1, or a pharmaceutically acceptable salt thereof, or apharmaceutical composition comprising the therapeutically effectiveamount of the compound or the pharmaceutically acceptable salt thereof,wherein the patient is a human.
 19. The method of claim 18, wherein thecancer is selected from the group consisting of: osteoma, hemangioma,granuloma, xanthoma, osteitis deformans, meningioma, meningiosarcoma,and gliomatosis, astrocytoma, medulloblastoma, glioma, ependymoma,germinoma (pinealoma), glioblastoma multiform, oligodendroglioma,oligodendrocytoma, schwannoma, retinoblastoma, and congenital tumors,neurofibroma, meningioma, glioma, and sarcoma.
 20. The method of claim18, wherein the disorder or disease is a neurologicalcondition/disorder/disease, wherein the neurologicalcondition/disorder/disease is selected from the group consisting of:Alzheimer's disease, amyotrophic lateral sclerosis (ALS), down syndrome,frontotemporal dementia (FTD), FTD with Parkinsonism-17 (FTDP-17),behavioural variant frontotemporal dementia (bvFTD), FTD in patientswith motor neuron disease (MND) (amyotrophic lateral sclerosis, alsocalled FTD-ALS), corticobasal degeneration (CBD) (also calledcorticobasal ganglionic degeneration), progressive supranuclear palsy,primary progressive aphasia (PPA), Prion Diseases, globular glialtauopathy (GGT), myotonic dystrophy type 1 (DM1) (also called Steinertdisease), myotonic dystrophy type 2 (DM2) (also called proximal myotonicmyopathy), Guam complex, argyrophilic grain disease, dementiapugilistica, post-encephalitic parkinsonism, lewy body dementia,Parkinson's disease, Pick's disease, and diseases with pronouncedneurodegeneration, autism, dementia, epilepsy, Huntington's disease,multiple sclerosis; diseases and disorders associated with acquiredbrain injury, chronic traumatic encephalopathy, traumatic brain injury,tumor, stroke, frontotemporal dementias, dementia with lewy bodies,prion diseases, multiple system atrophy, inclusion body myositis,degenerative myopathies, diabetic neuropathy, metabolic neuropathies,endocrine neuropathies, orthostatic hypotension, and Charcot-Marie-Toothdisease.
 21. The method of claim 18, wherein the disorder or disease isa neurological condition/disorder/disease associated with tau protein,amyloid, alpha-synuclein, Tar DNA-binding Protein of 43KDa (TDP-43),Prion protein PrP or fused in sarcoma (FUS) pathology.
 22. The method ofclaim 18, wherein the disorder or disease is a fibrotic disorder,wherein the fibrotic disorder is selected from the group consisting of:skin fibrosis; scleroderma; progressive systemic fibrosis; lungfibrosis; muscle fibrosis; kidney fibrosis; glomerulosclerosis;glomerulonephritis; hypertrophic scar formation; uterine fibrosis; renalfibrosis; cirrhosis of the liver, liver fibrosis; adhesions; chronicobstructive pulmonary disease; fibrosis following myocardial infarction;pulmonary fibrosis; fibrosis and scarring associated withdiffuse/interstitial lung disease; central nervous system fibrosis;fibrosis associated with proliferative vitreoretinopathy (PVR);restenosis; endometriosis; ischemic disease, and radiation fibrosis. 23.The method of claim 18, wherein the disorder or disease is chronicinflammation associated eye disorders, joint pain, arthritis(rheumatoid, osteo, psoriatic gout), cancers (colon, breast, lung,pancreas, and others), gastrointestinal disorders (ulcerative colitisand inflammatory bowel diseases), pulmonary disorders (chronicobstructive pulmonary disorder and asthma), allergies, skin disorders(atopic dermatitis and psoriasis), diabetes, pancreatitis, tendonitis,hepatitis, heart disease, myocarditis, stroke, lupus, and neurologicaldisorders, multiple sclerosis, Parkinson's and dementia, Alzheimer'sdisease.
 24. A method of reducing angiogenesis in a patient, the methodcomprising administering to the patient a therapeutically effectiveamount of a compound of claim 1, or a pharmaceutically acceptable salt,or a pharmaceutical composition.